Solid forms of an epidermal growth factor receptor kinase inhibitor

ABSTRACT

The present invention provides a solid form and compositions thereof, which are useful as an inhibitor of EGFR kinases and which exhibit desirable characteristics for the same.

CROSS REFERENCE TO RELATED CASES

The present application claims priority to U.S. provisional applicationSer. No. 61/611,376, filed Mar. 15, 2012, the entirety of which ishereby incorporated by reference.

FIELD OF THE INVENTION

The present invention provides solid forms of a compound useful asmutant-selective inhibitors of epidermal growth factor receptor (EGFR)kinase. The invention also provides pharmaceutically acceptablecompositions comprising solid forms of the present invention and methodsof using the compositions in the treatment of various disorders.

Protein tyrosine kinases are a class of enzymes that catalyze thetransfer of a phosphate group from ATP or GTP to a tyrosine residuelocated on a protein substrate. Receptor tyrosine kinases act totransmit signals from the outside of a cell to the inside by activatingsecondary messaging effectors via a phosphorylation event. A variety ofcellular processes are promoted by these signals, includingproliferation, carbohydrate utilization, protein synthesis,angiogenesis, cell growth, and cell survival.

There is strong precedent for involvement of the EGFR in human cancerbecause over 60% of all solid tumors overexpress at least one of theseproteins or their ligands. Overexpression of EGFR is commonly found inbreast, lung, head and neck, bladder tumors.

Activating mutations in the tyrosine kinase domain of EGFR have beenidentified in patients with non-small cell lung cancer (Lin, N. U.;Winer, E. P., Breast Cancer Res 6: 204-210, 2004). The reversibleinhibitors Tarceva (erlotinib) and Iressa (gefitinib) currently arefirst-line therapy for non-small cell lung cancer patients withactivating mutations. The most common activating mutations are L858R anddelE746-A750.

Additionally, in the majority of patients that relapse, acquired drugresistance, such as by mutation of gatekeeper residue T790M, has beendetected in at least half of such clinically resistant patients.Moreover, T790M may also be pre-existing; there may be an independent,oncogenic role for the T790M mutation. For example, there are patientswith the L858R/T790M mutation who never received gefitinib treatment. Inaddition, germline EGFR T790M mutations are linked with certain familiallung cancers.

Current drugs in development, including second-generation covalentinhibitors, such as BIBW2992, HKI-272 and PF-0299804, are effectiveagainst the T790M resistance mutation but exhibit dose-limitingtoxicities due to concurrent inhibition of WT EGFR. Accordingly, thereremains a need to find mutant-selective EGFR kinase inhibitors useful astherapeutic agents.

SUMMARY OF THE INVENTION

It has now been found that the novel solid forms of the presentinvention, and compositions thereof, are useful as mutant-selectiveinhibitors of one or more EGFR kinases and exhibits desirablecharacteristics for the same. In general, these solid forms, andpharmaceutically acceptable compositions thereof, are useful fortreating or lessening the severity of a variety of diseases or disordersas described in detail herein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts the x-ray powder diffraction (XRPD) pattern for Form A ofCompound 1.

FIG. 2 depicts the thermogravimetric analysis/differential thermalanalyser (TGA/DTA) pattern for Form A of Compound 1.

FIG. 3 depicts the differential scanning calorimetry (DSC) pattern forForm A of Compound 1.

FIG. 4 depicts the infrared (IR) spectrum for Form A of Compound 1.

FIG. 5 depicts the dynamic vapour sorption (DVS) pattern for Form A ofCompound 1.

FIG. 6 depicts the XRPD pattern for Form B of Compound 1.

FIG. 7 depicts the TGA/DTA pattern for Form B of Compound 1.

FIG. 8 depicts the DSC pattern for Form B of Compound 1.

FIG. 9 depicts the IR spectrum for Form B of Compound 1.

FIG. 10 depicts the DVS pattern for Form B of Compound 1.

FIG. 11 depicts the change in the XRPD pattern for Form B of Compound 1when heated to 120° C. and 160° C., respectively, as compared to Forms Aand B at ambient temperature.

FIG. 12 depicts the XRPD pattern for Form C of Compound 1.

FIG. 13 depicts the TGA/DTA pattern for Form C of Compound 1.

FIG. 14 depicts the DSC pattern for Form C of Compound 1.

FIG. 15 depicts the IR spectrum for Form C of Compound 1.

FIG. 16 depicts the DVS pattern for Form C of Compound 1.

FIG. 17 depicts the change in the XRPD pattern for Form C of Compound 1when stored at 40° C./75% RH for 1 week or 2 weeks, respectively, ascompared to the initial forms of Form A and Form C.

FIG. 18 depicts the change in the XRPD pattern for Form C of Compound 1when heated to 40° C. and 120° C., as compared to Forms A and C prior todrying.

FIG. 19 depicts the XRPD pattern for Form D of Compound 1, which ispresent in a mixture with Form B.

FIGS. 20A and B depict TGA/DTA patterns for Form D of Compound 1.

FIG. 21 depicts the DSC pattern for Form D of Compound 1.

FIG. 22 depicts the IR spectrum for Form D of Compound 1.

FIG. 23 depicts the DVS pattern for Form D of Compound 1.

FIG. 24 depicts the change in the XRPD pattern for Form D of Compound 1when stored at 40° C./75% RH for 1 week or 2 weeks, respectively, ascompared to the initial forms of Form A and Form D.

FIG. 25 depicts the XRPD pattern for Form E of Compound 1.

FIG. 26 depicts the TGA/DTA pattern for Form E of Compound 1.

FIG. 27 depicts the DSC pattern for Form E of Compound 1.

FIG. 28 depicts the IR spectrum for Form E of Compound 1.

FIG. 29 depicts the DVS pattern for Form E of Compound 1.

FIG. 30 depicts the change in the XRPD pattern for Form E of Compound 1when stored at 40° C./75% RH for 1 week or 2 weeks, respectively, ascompared to the initial forms of Form A and Form E.

FIG. 31 depicts the change in the XRPD pattern for Form E of Compound 1when heated to 40° C. and 120° C., as compared to Forms A and E prior todrying.

FIG. 32 depicts the XRPD pattern for Form F of Compound 1.

FIG. 33 depicts the TGA/DTA pattern for Form F of Compound 1.

FIG. 34 depicts the DSC pattern for Form F of Compound 1.

FIG. 35 depicts the IR spectrum for Form F of Compound 1.

FIG. 36 depicts the DVS pattern for Form F of Compound 1.

FIG. 37 depicts the change in the XRPD pattern for Form F of Compound 1when stored at 40° C./75% RH for 1 week or 2 weeks, respectively, ascompared to the initial forms of Form A and Form F.

FIG. 38 depicts the change in the XRPD pattern for Form F of Compound 1when heated to 40° C. and 120° C., as compared to Forms A and F prior todrying.

FIG. 39 depicts the XRPD pattern for Form G of Compound 1.

FIGS. 40A, 40B and 40C depict TGA/DTA patterns for Form G of Compound 1.

FIG. 41 depicts the DSC pattern for Form G of Compound 1.

FIG. 42 depicts the IR spectrum for Form G of Compound 1.

FIG. 43 depicts the DVS pattern for Form G of Compound 1.

FIG. 44 depicts the change in the XRPD pattern for Form G of Compound 1when stored at 40° C./75% RH for 1 week or 2 weeks, respectively, ascompared to the initial forms of Form A and Form G.

FIG. 45 depicts the change in the XRPD pattern for Form G of Compound 1when heated to 40° C. and 80° C., as compared to Forms A and G prior todrying.

FIG. 46 depicts the XRPD pattern for Form H of Compound 1.

FIG. 47 depicts the TGA/DTA pattern for Form H of Compound 1.

FIG. 48 depicts the XRPD pattern for Form H of Compound 1 after heatingto 115° C.

FIG. 49 depicts the DSC pattern for Form H of Compound 1.

FIG. 50 depicts the TGA/DTA pattern for Form H of Compound 1 when heatedto 115° C.

FIG. 51 depicts the TGA/DTA pattern for Form H of Compound 1 leaving theheated material on a bench for approximately one hour.

FIG. 52 depicts the XRPD for Form I of Compound 1.

FIG. 53 depicts the IR spectra for Form I of Compound 1.

FIG. 54 depicts the ¹H NMR spectrum for Form I of Compound 1.

FIG. 55 depicts the TGA/DTA thermogram for Form I of Compound 1following 3 days of drying under vacuum.

FIG. 56 depicts the TGA/DTA thermogram for Form I of Compound 1 afterdrying and standing at ambient for ca. 1 hour.

FIG. 57 depicts the DSC thermogram for Form I of Compound 1.

FIG. 58 depicts the DVS analysis for Form I of Compound 1.

FIG. 59 depicts the Post DVS XRPD analysis for Form I of Compound 1.

FIG. 60 depicts the HPLC analysis for Form I of Compound 1.

FIG. 61 depicts the XRPD analysis of solids remaining afterthermodynamic solubility studies of Form I of Compound 1.

FIG. 62 depicts the XRPD analysis of 1 week stability tests on Form I ofCompound 1 using open containers.

FIG. 63 depicts the XRPD analysis of 1 week stability tests on Form I ofCompound 1 using closed containers.

DETAILED DESCRIPTION OF THE INVENTION General Description of CertainAspects of the Invention

U.S. application Ser. No. 13/286,061 (“the '061 application”), filedOct. 31, 2011, the entirety of which is hereby incorporated herein byreference, describes certain 2,4-disubstituted pyrimidine compoundswhich covalently and irreversibly inhibit activity of EGFR kinase. Suchcompounds include Compound 1:

Compound 1(N-(3-(2-(4-(4-acetylpiperazin-1-yl)-2-methoxyphenylamino)-5-(trifluoromethyl)pyrimidin-4-ylamino)phenyl)acrylamide))is designated as compound number 1-4 and the synthesis of Compound 1 isdescribed in detail at Example 3 of the '061 application.

Compound 1 is active in a variety of assays and therapeutic modelsdemonstrating selective covalent, irreversible inhibition of mutant EGFRkinase (in enzymatic and cellular assays). Notably, Compound 1 was foundto inhibit human non-small cell lung cancer cell proliferation both invitro and in vivo. Accordingly, Compound 1 is useful for treating one ormore disorders associated with activity of mutant EGFR kinase.

It would be desirable to provide a solid form of Compound 1 that, ascompared to Compound 1, imparts characteristics such as improved aqueoussolubility, stability and ease of formulation. Accordingly, the presentinvention provides several solid forms of Compound 1.

According to one embodiment, the present invention provides an amorphousform, a crystalline form, or a mixture thereof. Exemplary solid formsare described in more detail below.

In other embodiments, the present invention provides Compound 1substantially free of impurities. As used herein, the term“substantially free of impurities” means that the compound contains nosignificant amount of extraneous matter. Such extraneous matter mayinclude starting materials, residual solvents, or any other impuritiesthat may result from the preparation of, and/or isolation of,Compound 1. In certain embodiments, at least about 90% by weight ofCompound 1 is present. In certain embodiments, at least about 95% byweight of Compound 1 is present. In still other embodiments of theinvention, at least about 99% by weight of Compound 1 is present.

According to one embodiment, Compound 1 is present in an amount of atleast about 95, 97, 97.5, 98.0, 98.5, 99, 99.5, 99.8 weight percentwhere the percentages are based on the total weight of the composition.According to another embodiment, Compound 1 contains no more than about5.0 area percent HPLC of total organic impurities and, in certainembodiments, no more than about 3.0 area percent HPLC of total organicimpurities and, in certain embodiments, no more than about 1.5 areapercent HPLC total organic impurities relative to the total area of theHPLC chromatogram. In other embodiments, Compound 1 contains no morethan about 1.0 area percent HPLC of any single impurity; no more thanabout 0.6 area percent HPLC of any single impurity, and, in certainembodiments, no more than about 0.5 area percent HPLC of any singleimpurity, relative to the total area of the HPLC chromatogram.

The structure depicted for Compound 1 is also meant to include alltautomeric forms of Compound 1. Additionally, structures depicted hereare also meant to include compounds that differ only in the presence ofone or more isotopically enriched atoms. For example, compounds havingthe present structure except for the replacement of hydrogen bydeuterium or tritium, or the replacement of a carbon by a ¹³C- or¹⁴C-enriched carbon are within the scope of this invention.

Solid Forms of Compound 1:

It has been found that Compound 1 can exist in a variety of solid forms.Such forms include polymorphs and amorphous forms. The solid forms canbe solvates, hydrates and unsolvated forms of Compound 1. All such formsare contemplated by the present invention. In certain embodiments, thepresent invention provides Compound 1 as a mixture of one or more solidforms of Compound 1.

As used herein, the term “polymorph” refers to the different crystalstructures (of solvated or unsolvated forms) in which a compound cancrystallize.

As used herein, the term “solvate” refers to a solid form with either astoichiometric or non-stoichiometric amount of solvent (e.g., a channelsolvate). For polymorphs, the solvent is incorporated into the crystalstructure. Similarly, the term “hydrate” refers to a solid form witheither a stoichiometric or non-stoichiometric amount of water. Forpolymorphs, the water is incorporated into the crystal structure.

As used herein, the term “about”, when used in reference to a degree2-theta value refers to the stated value ±0.3 degree 2-theta. In certainembodiments, “about” refers to ±0.2 degree 2-theta or ±0.1 degree2-theta.

In certain embodiments, Compound 1 is a crystalline solid. In otherembodiments, Compound 1 is a crystalline solid substantially free ofamorphous Compound 1. As used herein, the term “substantially free ofamorphous Compound 1” means that the compound contains no significantamount of amorphous Compound 1. In certain embodiments, at least about90% by weight of crystalline Compound 1 is present, or at least about95% by weight of crystalline Compound 1 is present. In still otherembodiments of the invention, at least about 97%, 98% or 99% by weightof crystalline compound 1 is present.

In certain embodiments, Compound 1 is a neat or unsolvated crystal formand thus does not have any water or solvent incorporated into thecrystal structure. It has been found that Compound 1 can exist in atleast two distinct neat (i.e., anhydrous) crystal forms, or polymorphs.In some embodiments, the present invention provides an anhydrouspolymorphic form of Compound 1 referred to herein as Form A. In otherembodiments, the present invention provides an anhydrous polymorphicform of Compound 1 referred to herein as Form B.

In certain embodiments, the present invention provides Form A ofCompound 1. According to one embodiment, Form A of Compound 1 ischaracterized by one or more peaks in its powder X-ray diffractionpattern selected from those at about 6.73, about 18.30, about 18.96 andabout 25.48 degrees 2-theta. In some embodiments, Form A of Compound 1is characterized by two or more peaks in its powder X-ray diffractionpattern selected from those at about 6.73, about 18.30, about 18.96 andabout 25.48 degrees 2-theta. In certain embodiments, Form A of Compound1 is characterized by three or more peaks in its powder X-raydiffraction pattern selected from those at about 6.73, about 18.30,about 18.96 and about 25.48 degrees 2-theta. In particular embodiments,Form A of Compound 1 is characterized by substantially all of the peaksin its X-ray powder diffraction pattern selected from those at about6.73, 14.24, 16.13, 18.30, 18.96, 20.59, 21.02, 21.23, 23.99 and 25.48degrees 2-theta. In an exemplary embodiment, Form A of Compound 1 ischaracterized by substantially all of the peaks in its X-ray powderdiffraction pattern selected from those at about

°2-Theta 5.21 5.42 6.73 8.67 9.47 10.59 10.93 12.07 13.00 13.06 13.6413.89 14.24 15.08 15.47 15.88 16.13 17.57 18.30 18.51 18.96 19.80 20.4320.59 21.02 21.23 22.18 22.93 23.99 24.22 25.48 26.18 26.50 27.68 30.3230.65 31.41 32.31 33.62 34.01 37.93 38.66 39.78 45.41

According to one aspect, Form A of Compound 1 has a powder X-raydiffraction pattern substantially similar to that depicted in FIG. 1.According to another aspect, Form A of Compound 1 has athermogravimetric analysis pattern substantially similar to thatdepicted in FIG. 2. Accordingly to yet another aspect, Form A ofCompound 1 has a differential scanning calorimetry pattern substantiallysimilar to that depicted in FIG. 3. According to a further embodiment,Form A of Compound 1 has a infrared spectrum substantially similar tothat depicted in FIG. 4. According to another embodiment, Form A ofCompound 1 has a dynamic vapour sorption pattern substantially similarto that depicted in FIG. 5. Form A of Compound 1 can be characterized bysubstantial similarity to two or more of these figures simultaneously.

In certain embodiments, the present invention provides Form B ofCompound 1. According to another embodiment, Form B of Compound 1 ischaracterized by one or more peaks in its powder X-ray diffractionpattern selected from those at about 10.67, about 12.21, about 18.11,about 19.24 and about 21.53 degrees 2-theta. In some embodiments, Form Bof Compound 1 is characterized by two or more peaks in its powder X-raydiffraction pattern selected from those at about 10.67, about 12.21,about 18.11, about 19.24 and about 21.53 degrees 2-theta. In certainembodiments, Form B of Compound 1 is characterized by three or morepeaks in its powder X-ray diffraction pattern selected from those atabout 10.67, about 12.21, about 18.11, about 19.24 and about 21.53degrees 2-theta. In particular embodiments, Form B of Compound 1 ischaracterized by substantially all of the peaks in its X-ray powderdiffraction pattern selected from those at about 8.96, 10.67, 12.21,14.56, 16.49, 17.74, 18.11, 19.24, 19.90, 21.53 and 23.93 degrees2-theta. In an exemplary embodiment, Form B of Compound 1 ischaracterized by substantially all of the peaks in its X-ray powderdiffraction pattern selected from those at about:

°2-Theta 3.03 4.74 5.01 6.91 7.59 8.23 8.96 10.67 12.21 13.31 14.2814.56 15.29 15.59 16.49 16.98 17.42 17.74 18.11 19.24 19.90 21.53 22.2523.93 24.63 24.87 25.30 26.34 27.66 29.31 30.57 31.21 32.39 32.58 33.4134.38 36.19 39.13 40.01 41.81 45.49 48.16

According to one aspect, Form B of Compound 1 has a powder X-raydiffraction pattern substantially similar to that depicted in FIG. 6.According to another aspect, Form B of Compound 1 has athermogravimetric analysis pattern substantially similar to thatdepicted in FIG. 7. Accordingly to yet another aspect, Form B ofCompound 1 has a differential scanning calorimetry pattern substantiallysimilar to that depicted in FIG. 8. According to a further embodiment,Form B of Compound 1 has an infrared spectrum substantially similar tothat depicted in FIG. 9. According to another embodiment, Form B ofCompound 1 has a dynamic vapour sorption pattern substantially similarto that depicted in FIG. 10. Form B of Compound 1 can be characterizedby substantial similarity to two or more of these figuressimultaneously.

In certain embodiments, Compound 1 is a dimethylformamide (DMF) solvatecrystal form. In some embodiments, the present invention provides a DMFsolvate polymorphic form of Compound 1 referred to herein as Form C.

In certain embodiments, the present invention provides Form C ofCompound 1. According one embodiment, Form C of Compound 1 ischaracterized by one or more peaks in its powder X-ray diffractionpattern selected from those at about 16.32, about 18.82, about 20.26,about 22.58 and about 25.36 degrees 2-theta. In some embodiments, Form Cof Compound 1 is characterized by two or more peaks in its powder X-raydiffraction pattern selected from those at about 16.32, about 18.82,about 20.26, about 22.58 and about 25.36 degrees 2-theta. In certainembodiments, Form C of Compound 1 is characterized by three or morepeaks in its powder X-ray diffraction pattern selected from those atabout 16.32, about 18.82, about 20.26, about 22.58 and about 25.36degrees 2-theta. In particular embodiments, Form C of Compound 1 ischaracterized by substantially all of the peaks in its X-ray powderdiffraction pattern selected from those at about 8.14, 14.45, 15.37,16.33, 18.16, 18.82, 20.26, 22.58, 22.96, 24.33, 25.36 and 26.36 degrees2-theta. In an exemplary embodiment, Form C of compound 1 ischaracterized by substantially all of the peaks in its X-ray powderdiffraction pattern selected from those at about:

°2-Theta 4.11 5.95 6.30 7.40 7.80 8.14 9.21 10.09 11.01 11.87 12.5713.59 14.45 15.37 15.94 16.33 16.67 17.03 17.57 18.16 18.37 18.82 19.3519.72 20.26 20.62 21.02 21.56 22.10 22.58 22.96 23.99 24.33 24.62 25.3626.36 27.02 27.37 27.81 28.44 29.12 29.45 29.80 30.28 30.66 31.24 31.7932.65 33.04 34.03 34.16 34.51 35.25 35.65 36.92 38.42 39.28 39.89 41.6442.14 44.15 44.54 45.35 46.02 46.44 48.42 49.16

According to one aspect, Form C of Compound 1 has a powder X-raydiffraction pattern substantially similar to that depicted in FIG. 12.According to another aspect, Form C of Compound 1 has athermogravimetric analysis pattern substantially similar to thatdepicted in FIG. 13. Accordingly to yet another aspect, Form C ofCompound 1 has a differential scanning calorimetry pattern substantiallysimilar to that depicted in FIG. 14. According to a further embodiment,Form C of Compound 1 has a infrared spectrum substantially similar tothat depicted in FIG. 15. According to another embodiment, Form C ofCompound 1 has a dynamic vapour sorption pattern substantially similarto that depicted in FIG. 16. Form C of Compound 1 can be characterizedby substantial similarity to two or more of these figuressimultaneously.

In certain embodiments, Compound 1 is a 1,4-dioxane solvate crystalform. In some embodiments, the present invention provides a 1,4-dioxanesolvate polymorphic form of Compound 1 referred to herein as Form D.

In certain embodiments, the present invention provides Form D ofCompound 1. According one embodiment, Form D of Compound 1 ischaracterized by one or more peaks in its powder X-ray diffractionpattern selected from those at about 18.40, about 19.31, about 20.14,about 20.53 and about 25.25 degrees 2-theta. In some embodiments, Form Dof Compound 1 is characterized by two or more peaks in its powder X-raydiffraction pattern selected from those at about 18.40, about 19.31,about 20.14, about 20.53 and about 25.25 degrees 2-theta. In certainembodiments, Form D of Compound 1 is characterized by three or morepeaks in its powder X-ray diffraction pattern selected from those atabout 18.40, about 19.31, about 20.14, about 20.53 and about 25.25degrees 2-theta. In particular embodiments, Form D of Compound 1 ischaracterized by substantially all of the peaks in its X-ray powderdiffraction pattern selected from those at about 13.51, 16.97, 17.86,18.40, 19.31, 20.14, 20.53, 21.04, 22.50, 24.98 and 25.25 degrees2-theta. In an exemplary embodiment, Form D of Compound 1 ischaracterized by substantially all of the peaks in its X-ray powderdiffraction pattern selected from those at about:

°2-Theta 3.03 3.30 5.49 6.60 6.86 7.21 7.56 8.93 9.87 10.11 10.68 11.3612.22 13.51 14.61 14.92 15.26 16.15 16.45 16.61 16.97 17.86 18.40 18.7719.31 20.14 20.53 21.04 21.52 22.50 23.80 24.98 25.25 25.64 26.35 26.6026.82 27.21 28.19 28.90 29.86

According to one aspect, Form D of Compound 1 has a powder X-raydiffraction pattern substantially similar to that depicted in FIG. 19.According to another aspect, Form D of Compound 1 has athermogravimetric analysis pattern substantially similar to thatdepicted in FIG. 20A or 20B. Accordingly to yet another aspect, Form Dof Compound 1 has a differential scanning calorimetry patternsubstantially similar to that depicted in FIG. 21. According to afurther embodiment, Form D of Compound 1 has an infrared spectrumsubstantially similar to that depicted in FIG. 22. According to anotherembodiment, Form D of Compound 1 has a dynamic vapour sorption patternsubstantially similar to that depicted in FIG. 23. Form D of Compound 1can be characterized by substantial similarity to two or more of thesefigures simultaneously.

In certain embodiments, Compound 1 is a methyl ethyl ketone (MEK)crystal form. In some embodiments, the present invention provides a MEKsolvate polymorphic form of Compound 1 referred to herein as Form E.

In certain embodiments, the present invention provides Form E ofCompound 1. According one embodiment, Form E of Compound 1 ischaracterized by one or more peaks in its powder X-ray diffractionpattern selected from those at about 5.78, about 12.57, about 15.34,about 19.10 and about 24.80 degrees 2-theta. In some embodiments, Form Eof Compound 1 is characterized by two or more peaks in its powder X-raydiffraction pattern selected from those at about 5.78, about 12.57,about 15.34, about 19.10 and about 24.80 degrees 2-theta. In certainembodiments, Form E of Compound 1 is characterized by three or morepeaks in its powder X-ray diffraction pattern selected from those atabout 5.78, about 12.57, about 15.34, about 19.10 and about 24.80degrees 2-theta. In particular embodiments, Form E of Compound 1 ischaracterized by substantially all of the peaks in its X-ray powderdiffraction pattern selected from those at about 5.78, 12.38, 12.57,14.14, 15.34, 18.22, 19.10, 20.05, 24.36 and 24.80 degrees 2-theta. Inan exemplary embodiment, Form E of Compound 1 is characterized bysubstantially all of the peaks in its X-ray powder diffraction patternselected from those at about:

°2-Theta 3.96 5.78 7.32 8.05 9.06 9.34 10.09 10.66 11.67 12.38 12.5713.92 14.14 15.34 15.66 16.28 16.84 17.56 17.97 18.22 18.84 19.10 20.0520.81 21.04 22.23 23.12 24.36 24.80 26.45 27.20 28.68 29.37 29.70 30.1330.70 32.37 33.06 33.43 34.37 35.75 39.30 41.13 47.08

According to one aspect, Form E of Compound 1 has a powder X-raydiffraction pattern substantially similar to that depicted in FIG. 25.According to another aspect, Form E of Compound 1 has athermogravimetric analysis pattern substantially similar to thatdepicted in FIG. 26. Accordingly to yet another aspect, Form E ofCompound 1 has a differential scanning calorimetry pattern substantiallysimilar to that depicted in FIG. 27. According to a further embodiment,Form E of Compound 1 has a infrared spectrum substantially similar tothat depicted in FIG. 28. According to another embodiment, Form E ofCompound 1 has a dynamic vapour sorption pattern substantially similarto that depicted in FIG. 29. Form E of Compound 1 can be characterizedby substantial similarity to two or more of these figuressimultaneously.

In certain embodiments, Compound 1 is a N-methyl-2-pyrrolidone (NMP)solvate crystal form. It has been found that Compound 1 can exist in atleast two distinct NMP crystal forms, or polymorphs. In someembodiments, the present invention provides a NMP solvate polymorphicform of Compound 1 referred to herein as Form F. In other embodiments,the present invention provides a NMP solvate polymorphic form ofCompound 1 referred to herein as Form G.

In certain embodiments, the present invention provides Form F ofCompound 1. According one embodiment, Form F of Compound 1 ischaracterized by one or more peaks in its powder X-ray diffractionpattern selected from those at about 15.51, about 16.86, about 18.80,about 20.97 and about 23.32 degrees 2-theta. In some embodiments, Form Fof Compound 1 is characterized by two or more peaks in its powder X-raydiffraction pattern selected from those at about 15.51, about 16.86,about 18.80, about 20.97 and about 23.32 degrees 2-theta. In certainembodiments, Form F of Compound 1 is characterized by three or morepeaks in its powder X-ray diffraction pattern selected from those atabout 15.51, about 16.86, about 18.80, about 20.97 and about 23.32degrees 2-theta. In particular embodiments, Form F of Compound 1 ischaracterized by substantially all of the peaks in its X-ray powderdiffraction pattern selected from those at about 5.64, 10.32, 12.97,13.54, 15.51, 16.39, 16.86, 18.80, 19.16, 20.97, 23.32 and 24.55 degrees2-theta. In an exemplary embodiment, Form F of Compound 1 ischaracterized by substantially all of the peaks in its X-ray powderdiffraction pattern selected from those at about:

°2-Theta 3.68 3.87 5.10 5.64 5.81 7.34 8.21 9.81 10.32 12.97 13.54 14.1215.00 15.51 16.39 16.86 17.36 18.80 19.16 19.68 20.08 20.97 21.93 22.6423.32 23.87 24.55 25.25 25.71 26.22 26.40 26.64 27.07 27.76 28.98 30.1130.95 31.16 31.46 34.04 34.56 35.72 36.70 37.60 38.68 39.55 40.14 40.8741.96 44.15 44.69 45.38 48.22

According to one aspect, Form F of Compound 1 has a powder X-raydiffraction pattern substantially similar to that depicted in FIG. 32.According to another aspect, Form F of Compound 1 has athermogravimetric analysis pattern substantially similar to thatdepicted in FIG. 33. Accordingly to yet another aspect, Form F ofCompound 1 has a differential scanning calorimetry pattern substantiallysimilar to that depicted in FIG. 34. According to a further embodiment,Form F of Compound 1 has a infrared spectrum substantially similar tothat depicted in FIG. 35. According to another embodiment, Form F ofCompound 1 has a dynamic vapour sorption pattern substantially similarto that depicted in FIG. 36. Form F of Compound 1 can be characterizedby substantial similarity to two or more of these figuressimultaneously.

In certain embodiments, the present invention provides Form G ofCompound 1. According to another embodiment, Form G of Compound 1 ischaracterized by one or more peaks in its powder X-ray diffractionpattern selected from those at about 6.79, about 17.86, about 19.43,about 19.98 and about 22.35 degrees 2-theta. In some embodiments, Form Gof Compound 1 is characterized by two or more peaks in its powder X-raydiffraction pattern selected from those at about 6.79, about 17.86,about 19.43, about 19.98 and about 22.35 degrees 2-theta. In furtherembodiments, Form G of Compound 1 is characterized by three or morepeaks in its powder X-ray diffraction pattern selected from those atabout 6.79, about 17.86, about 19.43, about 19.98 and about 22.35degrees 2-theta. In particular embodiments, Form G of Compound 1 ischaracterized by substantially all of the peaks in its X-ray powderdiffraction pattern selected from those at about 6.79, 6.89, 16.50,17.86, 19.43, 19.98, 22.35, 23.77 and 24.06 degrees 2-theta. In anexemplary embodiment, Form G of Compound 1 is characterized bysubstantially all of the peaks in its X-ray powder diffraction patternselected from those at about:

°2-Theta 3.70 4.38 6.79 6.89 8.60 9.85 12.28 13.48 14.52 15.35 16.0816.50 17.16 17.86 18.67 19.43 19.98 20.66 21.06 21.56 22.35 23.77 24.0625.03 26.06 26.22 26.79 27.56 28.07 28.74 29.18 29.59 30.08 30.43 31.2932.36 32.68 33.49 34.40 34.82 35.27 36.01 36.40 37.02 38.03 38.48 39.3739.94 41.00 41.97 42.64 43.69 44.91 45.35 46.20 47.43 48.53 48.75 49.48

According to one aspect, Form G of Compound 1 has a powder X-raydiffraction pattern substantially similar to that depicted in FIG. 39.According to another aspect, Form G of Compound 1 has athermogravimetric analysis pattern substantially similar to thatdepicted in any of FIG. 40A, 40B or 40C. Accordingly to yet anotheraspect, Form G of Compound 1 has a differential scanning calorimetrypattern substantially similar to that depicted in FIG. 41. According toa further embodiment, Form G of Compound 1 has an infrared spectrumsubstantially similar to that depicted in FIG. 42. According to anotherembodiment, Form G of Compound 1 has a dynamic vapour sorption patternsubstantially similar to that depicted in FIG. 43. Form G of Compound 1can be characterized by substantial similarity to two or more of thesefigures simultaneously.

In certain embodiments, Compound 1 is a hydrated crystal form. It hasbeen found that Compound 1 can exist in at least two distinct hydratedcrystal forms, or polymorphs. In some embodiments, the present inventionprovides a hydrated polymorphic form of Compound 1 referred to herein asForm H. In some embodiments, the present invention provides a hydratedpolymorphic form of Compound 1 referred to herein as Form I.

In certain embodiments, the present invention provides Form H ofCompound 1. According one embodiment, Form H of Compound 1 ischaracterized by one or more peaks in its powder X-ray diffractionpattern selected from those at about 10.82, about 11.08, about 18.45,about 22.85 and about 25.06 degrees 2-theta. In some embodiments, Form Hof Compound 1 is characterized by two or more peaks in its powder X-raydiffraction pattern selected from those at about 10.82, about 11.08,about 18.45, about 22.85 and about 25.06 degrees 2-theta. In certainembodiments, Form H of Compound 1 is characterized by three or morepeaks in its powder X-ray diffraction pattern selected from those atabout 10.82, about 11.08, about 18.45, about 22.85 and about 25.06degrees 2-theta. In particular embodiments, Form H of Compound 1 ischaracterized by substantially all of the peaks in its X-ray powderdiffraction pattern selected from those at about 10.14, 10.82, 11.08,18.45, 22.85, 24.33, 25.06 and 26.54 degrees 2-theta. In an exemplaryembodiment, Form H of Compound 1 is characterized by substantially allof the peaks in its X-ray powder diffraction pattern selected from thoseat about:

°2-Theta 3.56 5.34 6.42 8.57 8.82 9.79 10.14 10.82 11.08 11.92 13.6014.22 15.50 16.28 16.55 17.25 18.07 18.45 19.16 20.02 20.42 21.56 22.8523.31 24.33 25.06 25.91 26.02 26.54 27.36 27.45 27.79 28.31 29.15 29.45

According to one aspect, Form H of Compound 1 has a powder X-raydiffraction pattern substantially similar to that depicted in FIG. 46.According to another aspect, Form H of Compound 1 has athermogravimetric analysis pattern substantially similar to thatdepicted in FIG. 47. Accordingly to yet another aspect, Form H ofCompound 1 has a differential scanning calorimetry pattern substantiallysimilar to that depicted in FIG. 49. In some embodiments, Form H ofCompound 1 has a thermogravimetric analysis pattern substantiallysimilar to that depicted in FIG. 50 or FIG. 51. Form H of Compound 1 canbe characterized by substantial similarity to two or more of thesefigures simultaneously.

In certain embodiments, the present invention provides Form I ofCompound 1. According to one embodiment, Form I of Compound 1 ischaracterized by one or more peaks in its powder X-ray diffractionpattern selected from those at about 6.13, about 12.22, about 15.91,about 18.35, about 18.88, and about 21.90 degrees 2-theta. In someembodiments, Form I of Compound 1 is characterized by two or more peaksin its powder X-ray diffraction pattern selected from those at about6.13, about 12.22, about 15.91, about 18.35, about 18.88, and about21.90 degrees 2-theta. In some embodiments, Form I of Compound 1 ischaracterized by three or more peaks in its powder X-ray diffractionpattern selected from those at about 6.13, about 12.22, about 15.91,about 18.35, about 18.88, and about 21.90 degrees 2-theta. In someembodiments, Form I of Compound 1 is characterized by four or more peaksin its powder X-ray diffraction pattern selected from those at about6.13, about 12.22, about 15.91, about 18.35, about 18.88, and about21.90 degrees 2-theta. In some embodiments, Form I of Compound 1 ischaracterized by substantially all of the peaks in its powder X-raydiffraction pattern selected from those at about 6.13, about 12.22,about 15.91, about 18.35, about 18.88, and about 21.90 degrees 2-theta.In some embodiments, Form I of Compound 1 is characterized bysubstantially all of the peaks selected from those at about:

°2-Theta 5.39 6.13 7.65 9.33 10.18 12.22 12.72 12.98 14.56 15.08 15.3115.91 16.47 18.35 18.88 19.65 20.36 21.16 21.90 22.64 23.26 23.75 24.5524.95 25.78 27.95 28.92 29.47

According to one aspect, Form I of Compound 1 has a X-ray powderdiffraction pattern substantially similar to that depicted in FIG. 52.In some embodiments, Form I of Compound 1 has an infrared spectrumsubstantially similar to that depicted in FIG. 53. In some embodiments,Form I of Compound 1 has a ¹H NMR spectrum substantially similar to thatdepicted in FIG. 54. According to another aspect, Form I of Compound 1has a thermogravimetric analysis pattern substantially similar to thatdepicted in FIG. 55 or FIG. 56. Accordingly to yet another aspect, FormI of Compound 1 has a differential scanning calorimetry patternsubstantially similar to that depicted in FIG. 57. In some embodiments,Form I of Compound 1 has an dynamic vapour sorption substantiallysimilar to that depicted in FIG. 58. Form I of Compound 1 can becharacterized by substantial similarity to two or more of these figuressimultaneously.

It will be appreciated that any of the above-described polymorph formscan be characterized, for example, by reference to any of the peaks intheir respective X-ray diffraction patterns. Accordingly, in someembodiments, a polymorph described herein is characterized by one, two,three, four, five, six, seven, eight, nine, ten, eleven, twelve,thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen,twenty or more XRPD peaks (° 20). According to another embodiment, thepresent invention provides compound 1 as an amorphous solid. Amorphoussolids are well known to one of ordinary skill in the art and aretypically prepared by such methods as lyophilization, melting, andprecipitation from supercritical fluid, among others.

General Methods of Providing Compound 1:

Compound 1 is prepared according to the methods described in detail inthe '061 application, the entirety of which is hereby incorporatedherein by reference. The various solid forms of Compound 1 can beprepared by dissolving compound 1 in various suitable solvents and thencausing Compound 1 to return to the solid phase. Specific combinationsof solvents and conditions under which Compound 1 return to the solidphase are discussed in greater detail in the Examples.

A suitable solvent may solubilize Compound 1, either partially orcompletely. Examples of suitable solvents useful in the presentinvention are a protic solvent, a polar aprotic solvent, or mixturesthereof. In certain embodiments, suitable solvents include an ether, anester, an alcohol, a ketone, or a mixture thereof. In certainembodiments, the suitable solvent is methanol, ethanol, isopropanol, oracetone wherein said solvent is anhydrous or in combination with water,methyl tert-butyl ether (MTBE) or heptane. In other embodiments,suitable solvents include tetrahydrofuran, 1,4-dioxane,dimethylformamide, dimethylsulfoxide, glyme, diglyme, methyl ethylketone, N-methyl-2-pyrrolidone, methyl t-butyl ether, t-butanol,n-butanol, and acetonitrile. In another embodiment, the suitable solventis anhydrous ethanol. In some embodiments, the suitable solvent is MTBE.

According to another embodiment, the present invention provides a methodfor preparing a solid form of Compound 1, comprising the steps ofdissolving Compound 1 with a suitable solvent and optionally heating toform a solution thereof; and isolating Compound 1.

As described generally above, Compound 1 is dissolved in a suitablesolvent, optionally with heating. In certain embodiments Compound 1 isdissolved at about 50 to about 60° C. In other embodiments, Compound 1is dissolved at about 50 to about 55° C. In still other embodiments,Compound 1 is dissolved at the boiling temperature of the solvent. Inother embodiments, Compound 1 is dissolved without heating (e.g., atambient temperature, approximately 20-25° C.).

In certain embodiments, Compound 1 precipitates from the mixture. Inanother embodiment, Compound 1 crystallizes from the mixture. In otherembodiments, Compound 1 crystallizes from solution following seeding ofthe solution (i.e., adding crystals of Compound 1 to the solution).

Crystalline Compound 1 can precipitate out of the reaction mixture, orbe generated by removal of part or all of the solvent through methodssuch as evaporation, distillation, filtration (e.g., nanofiltration,ultrafiltration), reverse osmosis, absorption and reaction, by adding ananti-solvent (e.g., water, MTBE and/or heptane), by cooling (e.g., crashcooling) or by different combinations of these methods.

As described generally above, Compound 1 is optionally isolated. It willbe appreciated that Compound 1 may be isolated by any suitable physicalmeans known to one of ordinary skill in the art. In certain embodiments,precipitated solid Compound 1 is separated from the supernatant byfiltration. In other embodiments, precipitated solid Compound 1 isseparated from the supernatant by decanting the supernatant.

In certain embodiments, precipitated solid Compound 1 is separated fromthe supernatant by filtration.

In certain embodiments, isolated Compound 1 is dried in air. In otherembodiments isolated Compound 1 is dried under reduced pressure,optionally at elevated temperature.

Uses, Formulation and Administration Pharmaceutically AcceptableCompositions

According to another embodiment, the invention provides a compositioncomprising Compound 1 and a pharmaceutically acceptable carrier,adjuvant, or vehicle. The amount of Compound 1 in compositions of thisinvention is such that it is effective to measurably inhibit a proteinkinase, particularly an EGFR kinase, or a mutant thereof, in abiological sample or in a patient. In certain embodiments, a compositionof this invention is formulated for administration to a patient in needof such composition. In some embodiments, a composition of thisinvention is formulated for oral administration to a patient.

The term “patient”, as used herein, means an animal, preferably amammal, and most preferably a human.

The term “pharmaceutically acceptable carrier, adjuvant, or vehicle”refers to a non-toxic carrier, adjuvant, or vehicle that does notdestroy the pharmacological activity of the compound with which it isformulated. Pharmaceutically acceptable carriers, adjuvants or vehiclesthat may be used in the compositions of this invention include, but arenot limited to, ion exchangers, alumina, aluminum stearate, lecithin,serum proteins, such as human serum albumin, buffer substances such asphosphates, glycine, sorbic acid, potassium sorbate, partial glyceridemixtures of saturated vegetable fatty acids, water, salts orelectrolytes, such as protamine sulfate, disodium hydrogen phosphate,potassium hydrogen phosphate, sodium chloride, zinc salts, colloidalsilica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-basedsubstances, polyethylene glycol, Vitamin E polyethylene glycol succinate(d-alpha tocopheryl polyethylene glycol 1000 succinate), sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, and wool fat.

Compositions of the present invention may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal,intrahepatic, intralesional and intracranial injection or infusiontechniques. Preferably, the compositions are administered orally,intraperitoneally or intravenously. Sterile injectable forms of thecompositions of this invention may be an aqueous or oleaginoussuspension. These suspensions may be formulated according to techniquesknown in the art using suitable dispersing or wetting agents andsuspending agents. The sterile injectable preparation may also be asterile injectable solution or suspension in a non-toxic parenterallyacceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium.

For this purpose, any bland fixed oil may be employed includingsynthetic mono- or di-glycerides. Fatty acids, such as oleic acid andits glyceride derivatives are useful in the preparation of injectables,as are natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant, such as carboxymethyl cellulose or similar dispersingagents that are commonly used in the formulation of pharmaceuticallyacceptable dosage forms including emulsions and suspensions. Othercommonly used surfactants, such as Tweens, Spans and other emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms may also be used for the purposes of formulation.

Pharmaceutically acceptable compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous and non-aqueous suspensions orsolutions. In the case of tablets for oral use, carriers commonly usedinclude lactose and corn starch. Lubricating agents, such as magnesiumstearate, are also typically added. For oral administration in a capsuleform, useful diluents include lactose and dried cornstarch. When aqueoussuspensions are required for oral use, the active ingredient istypically combined with emulsifying and suspending agents. If desired,certain sweetening, flavoring or coloring agents may also be added.

Alternatively, pharmaceutically acceptable compositions of thisinvention may be administered in the form of suppositories for rectaladministration. These can be prepared by mixing the agent with asuitable non-irritating excipient that is solid at room temperature butliquid at rectal temperature and therefore will melt in the rectum torelease the drug. Such materials include cocoa butter, beeswax andpolyethylene glycols.

Pharmaceutically acceptable compositions of this invention may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, provided pharmaceutically acceptablecompositions may be formulated in a suitable ointment containing theactive component suspended or dissolved in one or more carriers.Carriers for topical administration of Compound 1 include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, provided pharmaceutically acceptable compositionscan be formulated in a suitable lotion or cream containing the activecomponents suspended or dissolved in one or more pharmaceuticallyacceptable carriers. Suitable carriers include, but are not limited to,mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax,cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, provided pharmaceutically acceptable compositionsmay be formulated as micronized suspensions in isotonic, pH adjustedsterile saline, or, preferably, as solutions in isotonic, pH adjustedsterile saline, either with or without a preservative such asbenzylalkonium chloride. Alternatively, for ophthalmic uses, thepharmaceutically acceptable compositions may be formulated in anointment such as petrolatum.

Pharmaceutically acceptable compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

In some embodiments, pharmaceutically acceptable compositions of thisinvention are formulated for oral administration.

The amount of Compound 1 that may be combined with the carrier materialsto produce a composition in a single dosage form will vary dependingupon the host treated, the particular mode of administration. In certainembodiments, provided compositions are formulated so that a dosage ofbetween 0.01-100 mg/kg body weight/day of Compound 1 can be administeredto a patient receiving these compositions.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease being treated.

Uses of Compounds and Pharmaceutically Acceptable Compositions

Compound 1 and compositions described herein are generally useful forthe inhibition of protein kinase activity of one or more enzymes.Examples of kinases that are inhibited by Compound 1 and compositionsdescribed herein and against which the methods described herein areuseful include EGFR kinase or a mutant thereof. It has been found thatCompound 1 is a selective inhibitor of at least one mutation of EGFR, ascompared to wild-type (“WT”) EGFR. In certain embodiments, an at leastone mutation of EGFR is T790M. In certain embodiments, the at least onemutation of EGFR is a deletion mutation. In some embodiments, the atleast one mutation of EGFR is an activating mutation. In certainembodiments, Compound 1 selectively inhibits at least one resistantmutation and at least one activating mutation as compared to WT EGFR. Insome embodiments, Compound 1 selectively inhibits at least one deletionmutation and/or at least one point mutation, and is sparing as to WTEGFR inhibition.

A mutation of EGFR can be selected from T790M (resistant or oncogenic),L858R (activating), delE746-A750 (activating), G719S (activating), or acombination thereof.

As used herein, the term “selectively inhibits,” as used in comparisonto inhibition of WT EGFR, means that Compound 1 inhibits at least onemutation of EGFR (i.e., at least one deletion mutation, at least oneactivating mutation, at least one restistant mutation, or a combinationof at least one deletion mutation and at least one point mutation) in atleast one assay described herein (e.g., biochemical or cellular). Insome embodiments, the term “selectively inhibits,” as used in comparisonto WT EGFR inhibition means that Compound 1 is at least 50 times morepotent, at least 45 times, at least 40, at least 35, at least 30, atleast 25, or at least 20 times more potent as an inhibitor of at leastone mutation of EGFR, as defined and described herein, as compared to WTEGFR.

As used herein, the term “sparing as to WT EGFR” means that a selectiveinhibitor of at least one mutation of EGFR, as defined and describedabove and herein, inhibits EGFR at the upper limit of detection of atleast one assay, such as those described in the '061 application (e.g.,biochemical or cellular as described in detail in Examples 56-58). Invitro assays include assays that determine inhibition of thephosphorylation activity and/or the subsequent functional consequences,or ATPase activity of activated EGFR (WT or mutant). Alternate in vitroassays quantitate the ability of the inhibitor to bind to EGFR (WT ormutant). Inhibitor binding may be measured by radiolabeling theinhibitor prior to binding, isolating the inhibitor/EGFR (WT or mutant)complex and determining the amount of radiolabel bound. Alternatively,inhibitor binding may be determined by running a competition experimentwhere new inhibitors are incubated with EGFR (WT or mutant) bound toknown radioligands. In some embodiments, the term “sparing as to WTEGFR” means that Compound 1 inhibits WT EGFR with an IC₅₀ of at least 10μM, at least 9 μM, at least 8 μM, at least 7 μM, at least 6 μM, at least5 μM, at least 3 μM, at least 2 μM, or at least 1 μM.

In certain embodiments, Compound 1 selectively inhibits (a) at least oneactivating mutation; and (b) T790M; and (c) is sparing as to WT. In someembodiments, an at least one activating mutation is a deletion mutation.In some embodiments, an at least one activating mutation is a pointmutation. In some embodiments, an activating mutation is delE746-A750.In some embodiments, an activating mutation is L858R. In someembodiments, an activating mutation is G719S.

In some embodiments, the at least one mutation of EGFR is L858R and/orT790M.

Without wishing to be bound by any particular theory, it is believedthat administration of Compound 1 to a patient having at least oneactivating mutation may preempt formation of the T790M resistancemutation. Thus, in certain embodiments, the present invention provides amethod for inhibiting an activating mutation in a patient comprisingadministering to the patient Compound 1 or composition thereof, asdescribed herein.

One of ordinary skill in the art will appreciate that certain patientshave an oncogenic form of the T790M mutation, i.e., the T790M mutationis present prior to administrating any EGFR kinase inhibitor to apatient and is therefore oncogenic. Accordingly, in some embodiments,the present invention provides a method for inhibiting oncogenic T790Min a patient comprising administering to the patient a provided compoundor composition thereof, as described herein.

In certain embodiments, the amount of Compound 1 in a composition iseffective to measurably inhibit at least one mutant of EGFR selectivelyas compared to WT EGFR and other protein kinases (e.g., ErbB2, ErbB4, aTEC-kinase, and/or JAK3), in a biological sample or in a patient.

As used herein, the terms “treatment,” “treat,” and “treating” refer toreversing, alleviating, delaying the onset of, or inhibiting theprogress of a disease or disorder, or one or more symptoms thereof, asdescribed herein. In some embodiments, treatment may be administeredafter one or more symptoms have developed. In other embodiments,treatment may be administered in the absence of symptoms. For example,treatment may be administered to a susceptible individual prior to theonset of symptoms (e.g., in light of a history of symptoms and/or inlight of genetic or other susceptibility factors). Treatment may also becontinued after symptoms have resolved, for example to prevent or delaytheir recurrence.

Compound 1 is an inhibitor of at least one mutant of EGFR and istherefore useful for treating one or more disorders associated withactivity of one of more EGFR mutants (e.g., a deletion mutation, anactivating mutation, a resistant mutation, or combination thereof).Thus, in certain embodiments, the present invention provides a methodfor treating a mutant EGFR-mediated disorder comprising the step ofadministering to a patient in need thereof. Compound 1, orpharmaceutically acceptable composition thereof.

As used herein, the term “mutant EGFR-mediated” disorders or conditionsas used herein means any disease or other deleterious condition in whichat least one mutant of EGFR is known to play a role. In certainembodiments, an at least one mutant of EGFR is T790M. In someembodiments, the at least one mutant of EGFR is a deletion mutation. Incertain embodiments, the at least one mutant of EGFR is an activatingmutation. In some embodiments, the at least one mutant of EGFR is L858Rand/or T790M. In certain embodiments, Compound 1 selectively inhibits(a) at least one activating mutation, (b) T790M, and (c) is sparing asto WT. In some embodiments, an at least one activating mutation is adeletion mutation. In some embodiments, an at least one activatingmutation is a point mutation. In some embodiments, an activatingmutation is delE746-A750. In some embodiments, an activating mutation isL858R. In some embodiments, an activating mutation is G719S.

Accordingly, another embodiment of the present invention relates totreating or lessening the severity of one or more diseases in which atleast one mutant of EGFR is known to play a role. Specifically, thepresent invention relates to a method of treating or lessening theseverity of a disease or condition selected from a proliferativedisorder, wherein said method comprises administering to a patient inneed thereof a compound or composition according to the presentinvention.

In some embodiments, the present invention provides a method fortreating or lessening the severity of one or more disorders selectedfrom a cancer. In some embodiments, the cancer is associated with asolid tumor. In certain embodiments, the cancer is breast cancer,glioblastoma, lung cancer, cancer of the head and neck, colorectalcancer, bladder cancer, or non-small cell lung cancer. In someembodiments, the present invention provides a method for treating orlessening the severity of one or more disorders selected from squamouscell carcinoma, salivary gland carcinoma, ovarian carcinoma, orpancreatic cancer.

In certain embodiments, the present invention provides a method fortreating or lessening the severity of neurofibromatosis type I (NF1),neurofibromatosis type II (NF2) Schwann cell neoplasms (e.g. MPNST's),or Schwannomas.

Compound 1 and compositions thereof, according to the method of thepresent invention, may be administered using any amount and any route ofadministration effective for treating or lessening the severity of acancer. The exact amount required will vary from subject to subject,depending on the species, age, and general condition of the subject, theseverity of the infection, the particular agent, its mode ofadministration, and the like. Compound 1 is preferably formulated indosage unit form for ease of administration and uniformity of dosage.The expression “dosage unit form” as used herein refers to a physicallydiscrete unit of agent appropriate for the patient to be treated. Itwill be understood, however, that the total daily usage of the compoundsand compositions of the present invention will be decided by theattending physician within the scope of sound medical judgment. Thespecific effective dose level for any particular patient or organismwill depend upon a variety of factors including the disorder beingtreated and the severity of the disorder; the activity of the specificcompound employed; the specific composition employed; the age, bodyweight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed, andlike factors well known in the medical arts.

Pharmaceutically acceptable compositions of this invention can beadministered to humans and other animals orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), bucally, as an oral or nasal spray, orthe like, depending on the severity of the infection being treated. Incertain embodiments, Compound 1 may be administered orally orparenterally at dosage levels of about 0.01 mg/kg to about 50 mg/kg andpreferably from about 0.5 mg/kg to about 25 mg/kg, of subject bodyweight per day, one or more times a day, to obtain the desiredtherapeutic effect.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to Compound 1, the liquiddosage forms may contain inert diluents commonly used in the art suchas, for example, water or other solvents, solubilizing agents andemulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate,ethyl acetate, benzyl alcohol, benzyl benzoate, polyethylene glycol(e.g., PEG 200, PEG 400, PEG 1000, PEG 2000), propylene glycol,1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed,groundnut, corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, Vitamin E polyethylene glycol succinate(d-alpha tocopheryl polyethylene glycol 1000 succinate), polyethyleneglycols and fatty acid esters of sorbitan, and mixtures thereof. Besidesinert diluents, the oral compositions can also include adjuvants such aswetting agents, emulsifying and suspending agents, sweetening,flavoring, and perfuming agents. The liquid forms above can also befilled into a soft or hard capsule to form a solid dosage form. Suitablecapsules can be formed from, for example, gelatin, strach and cellulosederivatives (e.g., hydroxycellulose, hydropropylmethylcellulose).

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

Injectable formulations can be sterilized, for example, by filtrationthrough a bacterial-retaining filter, or by incorporating sterilizingagents in the form of sterile solid compositions which can be dissolvedor dispersed in sterile water or other sterile injectable medium priorto use.

In order to prolong the effect of Compound 1 of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, delayed absorption ofa parenterally administered compound form is accomplished by dissolvingor suspending the compound in an oil vehicle. Injectable depot forms aremade by forming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing Compound 1 of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, Compound 1 ismixed with at least one inert, pharmaceutically acceptable excipient orcarrier such as sodium citrate or dicalcium phosphate and/or a) fillersor extenders such as starches, lactose, sucrose, glucose, mannitol, andsilicic acid, b) binders such as, for example, carboxymethylcellulose,alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c)humectants such as glycerol, d) disintegrating agents such as agar-agar,calcium carbonate, potato or tapioca starch, alginic acid, certainsilicates, and sodium carbonate, e) solution retarding agents such asparaffin, f) absorption accelerators such as quaternary ammoniumcompounds, g) wetting agents such as, for example, cetyl alcohol andglycerol monostearate, h) absorbents such as kaolin and bentonite clay,and i) lubricants such as talc, calcium stearate, magnesium stearate,solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof.In the case of capsules, tablets and pills, the dosage form may alsocomprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled capsules using such excipients as lactose or milksugar as well as high molecular weight polyethylene glycols and thelike. The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled capsulesusing such excipients as lactose or milk sugar as well as high molecularweight polyethylene glycols and the like.

Compound 1 can also be in micro-encapsulated form with one or moreexcipients as noted above. The solid dosage forms of tablets, dragees,capsules, pills, and granules can be prepared with coatings and shellssuch as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of Compound 1include ointments, pastes, creams, lotions, gels, powders, solutions,sprays, inhalants or patches. The active component is admixed understerile conditions with a pharmaceutically acceptable carrier and anyneeded preservatives or buffers as may be required. Ophthalmicformulation, ear drops, and eye drops are also contemplated as beingwithin the scope of this invention. Additionally, the present inventioncontemplates the use of transdermal patches, which have the addedadvantage of providing controlled delivery of a compound to the body.Such dosage forms can be made by dissolving or dispensing the compoundin the proper medium. Absorption enhancers can also be used to increasethe flux of the compound across the skin. The rate can be controlled byeither providing a rate controlling membrane or by dispersing thecompound in a polymer matrix or gel.

According to another embodiment, the invention relates to a method ofinhibiting at least one mutant of EGFR (e.g., a deletion mutation, anactivating mutation, a resistant mutations, or combination thereof)activity in a biological sample comprising the step of contacting saidbiological sample with Compound 1, or a composition comprising thecompound. In certain embodiments, the invention relates to a method ofirreversibly inhibiting at least one mutant of EGFR (e.g., a deletionmutation, an activating mutation, a resistant mutation, or combinationthereof) activity in a biological sample comprising the step ofcontacting the biological sample with Compound 1, or a compositioncomprising the compound.

In certain embodiments, Compound 1 selectively inhibits in a biologicalsample (a) at least one activating mutation, (b) T790M, and (c) issparing as to WT. In some embodiments, an at least one activatingmutation is a deletion mutation. In some embodiments, an at least oneactivating mutation is a point mutation. In some embodiments, anactivating mutation is delE746-A750. In some embodiments, an activatingmutation is L858R. In some embodiments, an activating mutation is G719S.

The term “biological sample”, as used herein, includes, withoutlimitation, cell cultures or extracts thereof; biopsied materialobtained from a mammal or extracts thereof; and blood, saliva, urine,feces, semen, tears, or other body fluids or extracts thereof.

Inhibition of at least one mutant of EGFR (e.g., a deletion mutation, anactivating mutation, a resistant mutation, or combination thereof)activity in a biological sample is useful for a variety of purposes thatare known to one of skill in the art. Examples of such purposes include,but are not limited to, blood transfusion, organ transplantation,biological specimen storage, and biological assays.

Another embodiment of the present invention relates to a method ofinhibiting at least one mutant of EGFR (e.g., a deletion mutation, anactivating mutation, a resistant mutation, or combination thereof)activity in a patient comprising the step of administering to thepatient Compound 1 or a composition comprising the compound. In certainembodiments, the present invention provides a method for inhibiting (a)at least one activating mutation, and (b) T790M in a patient, and (c) issparing as to WT, wherein the method comprises administering to thepatient Compound 1 or composition thereof. In some embodiments, an atleast one activating mutation is a deletion mutation. In someembodiments, an at least one activating mutation is a point mutation. Insome embodiments, the present invention provides a method for inhibitingat least one mutant of EGFR in a patient, wherein an activating mutationis delE746-A750. In some embodiments, the present invention provides amethod for inhibiting at least one mutant of EGFR in a patient, whereinan activating mutation is L858R. In some embodiments, the presentinvention provides a method for inhibiting at least one mutant of EGFRin a patient, wherein an activating mutation is G719S.

According to another embodiment, the invention relates to a method ofinhibiting at least one mutant of EGFR (e.g., a deletion mutation, anactivating mutation, a resistant mutation, or combination thereof)activity in a patient comprising the step of administering to thepatient Compound 1 or a composition comprising the compound. Accordingto certain embodiments, the invention relates to a method ofirreversibly inhibiting at least one mutant of EGFR activity (e.g., adeletion mutation, an activating mutation, a resistant mutation, orcombination thereof) in a patient comprising the step of administeringto said patient Compound 1 or a composition comprising the compound. Incertain embodiments, the present invention provides a method forirreversibly inhibiting (a) at least one activating mutation, and (b)T790M in a patient, and (c) is sparing as to WT, wherein said methodcomprises administering to the patient Compound 1 or compositionthereof. In some embodiments, an irreversibly inhibited at least oneactivating mutation is a deletion mutation. In some embodiments, anirreversibly inhibited at least one activating mutation is a pointmutation. In some embodiments, the present invention provides a methodfor irreversibly inhibiting at least one mutant of EGFR in a patient,wherein an activating mutation is delE746-A750. In some embodiments, thepresent invention provides a method for irreversibly inhibiting at leastone mutant of EGFR in a patient, wherein an activating mutation isL858R. In some embodiments, the present invention provides a method forirreversibly inhibiting at least one mutant of EGFR in a patient,wherein an activating mutation is G719S.

In other embodiments, the present invention provides a method fortreating a disorder mediated by one or more of at least one mutant ofEGFR (e.g., a deletion mutation, an activating mutation, a resistantmutation, or combination thereof) in a patient in need thereof,comprising the step of administering to said patient Compound 1 orpharmaceutically acceptable composition thereof. Such disorders aredescribed in detail herein.

Depending upon the particular condition, or disease, to be treated,additional therapeutic agents, which are normally administered to treatthat condition, may also be present in the compositions of thisinvention or as part of a treatment regimen including Compound 1. Asused herein, additional therapeutic agents that are normallyadministered to treat a particular disease, or condition, are known as“appropriate for the disease or condition being treated.”

For example, Compound 1 or a pharmaceutically acceptable compositionthereof is administered in combination with chemotherapeutic agents totreat proliferative diseases and cancer. Examples of knownchemotherapeutic agents include, but are not limited to, Adriamycin,dexamethasone, vincristine, cyclophosphamide, fluorouracil, topotecan,taxol, interferons, platinum derivatives, taxane (e.g., paclitaxel),vinca alkaloids (e.g., vinblastine), anthracyclines (e.g., doxorubicin),epipodophyllotoxins (e.g., etoposide), cisplatin, an mTOR inhibitor(e.g., a rapamycin), methotrexate, actinomycin D, dolastatin 10,colchicine, emetine, trimetrexate, metoprine, cyclosporine,daunorubicin, teniposide, amphotericin, alkylating agents (e.g.,chlorambucil), 5-fluorouracil, camptothecin, cisplatin, metronidazole,and Gleevec™, among others. In other embodiments, Compound 1 isadministered in combination with a biologic agent, such as Avastin orVECTIBIX.

In certain embodiments, Compound 1 or a pharmaceutically acceptablecomposition thereof is administered in combination with anantiproliferative or chemotherapeutic agent selected from any one ormore of abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol,altretamine, amifostine, anastrozole, arsenic trioxide, asparaginase,azacitidine, BCG Live, bevacuzimab, fluorouracil, bexarotene, bleomycin,bortezomib, busulfan, calusterone, capecitabine, camptothecin,carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cladribine,clofarabine, cyclophosphamide, cytarabine, dactinomycin, darbepoetinalfa, daunorubicin, denileukin, dexrazoxane, docetaxel, doxorubicin(neutral), doxorubicin hydrochloride, dromostanolone propionate,epirubicin, epoetin alfa, erlotinib, estramustine, etoposide phosphate,etoposide, exemestane, filgrastim, floxuridine fludarabine, fulvestrant,gefitinib, gemcitabine, gemtuzumab, goserelin acetate, histrelinacetate, hydroxyurea, ibritumomab, idarubicin, ifosfamide, imatinibmesylate, interferon alfa-2a, interferon alfa-2b, irinotecan,lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole,lomustine, megestrol acetate, melphalan, mercaptopurine, 6-MP, mesna,methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone,nandrolone, nelarabine, nofetumomab, oprelvekin, oxaliplatin,paclitaxel, palifermin, pamidronate, pegademase, pegaspargase,pegfilgrastim, pemetrexed disodium, pentostatin, pipobroman, plicamycin,porfimer sodium, procarbazine, quinacrine, rasburicase, rituximab,sargramostim, sorafenib, streptozocin, sunitinib maleate, talc,tamoxifen, temozolomide, teniposide, VM-26, testolactone, thioguanine,6-TG, thiotepa, topotecan, toremifene, tositumomab, trastuzumab,tretinoin, ATRA, uracil mustard, valrubicin, vinblastine, vincristine,vinorelbine, zoledronate, or zoledronic acid.

Other examples of agents the inhibitors of this invention may also becombined with include, without limitation: treatments for Alzheimer'sDisease such as donepezil hydrochloride (Aricept®) and rivastigmine(Exelon®); treatments for Parkinson's Disease such as L-DOPA/carbidopa,entacapone, ropinrole, pramipexole, bromocriptine, pergolide,trihexephendyl, and amantadine; agents for treating Multiple Sclerosis(MS) such as beta interferon (e.g., Avonex® and Rebif®), glatirameracetate (Copaxone®), and mitoxantrone; treatments for asthma such asalbuterol and montelukast (Singulair®); agents for treatingschizophrenia such as zyprexa, risperdal, seroquel, and haloperidol;anti-inflammatory agents such as corticosteroids, TNF blockers, IL-1 RA,azathioprine, cyclophosphamide, and sulfasalazine; immunomodulatory andimmunosuppressive agents such as cyclosporin, tacrolimus, rapamycin,mycophenolate mofetil, interferons, corticosteroids, cyclophophamide,azathioprine, and sulfasalazine; neurotrophic factors such asacetylcholinesterase inhibitors, MAO inhibitors, interferons,anti-convulsants, ion channel blockers, riluzole, and anti-Parkinsonianagents; agents for treating cardiovascular disease such asbeta-blockers, ACE inhibitors, diuretics, nitrates, calcium channelblockers, and statins; agents for treating liver disease such ascorticosteroids, cholestyramine, interferons, and anti-viral agents;agents for treating blood disorders such as corticosteroids,anti-leukemic agents, and growth factors; and agents for treatingimmunodeficiency disorders such as gamma globulin.

In certain embodiments, Compound 1 or a pharmaceutically acceptablecomposition thereof is administered in combination with a monoclonalantibody or an siRNA therapeutic.

The additional agents may be administered separately from a Compound1-containing composition, as part of a multiple dosage regimen.Alternatively, those agents may be part of a single dosage form, mixedtogether with Compound 1 in a single composition. If administered aspart of a multiple dosage regime, the two active agents may be submittedsimultaneously, sequentially or within a period of time from one another(e.g., one hour, two hours, six hours, twelve hours, one day, one week,two weeks, one month).

As used herein, the terms “combination,” “combined,” and related termsrefer to the simultaneous or sequential administration of therapeuticagents in accordance with this invention. For example, Compound 1 may beadministered with another therapeutic agent simultaneously orsequentially in separate unit dosage forms or together in a single unitdosage form. Accordingly, the present invention provides a single unitdosage form comprising Compound 1, an additional therapeutic agent, anda pharmaceutically acceptable carrier, adjuvant, or vehicle.

The amount of Compound 1 and additional therapeutic agent (in thosecompositions which comprise an additional therapeutic agent as describedabove) that may be combined with the carrier materials to produce asingle dosage form will vary depending upon the host treated and theparticular mode of administration. Preferably, compositions of thisinvention should be formulated so that a dosage of between 0.01-100mg/kg body weight/day of Compound 1 can be administered.

In those compositions that include an additional therapeutic agent, thatadditional therapeutic agent and Compound 1 may act synergistically.Therefore, the amount of additional therapeutic agent in suchcompositions may be less than that required in a monotherapy utilizingonly that therapeutic agent. In such compositions, a dosage of between0.01-1,000 μg/kg body weight/day of the additional therapeutic agent canbe administered.

The amount of additional therapeutic agent present in the compositionsof this invention will be no more than the amount that would normally beadministered in a composition comprising that therapeutic agent as theonly active agent. Preferably the amount of additional therapeutic agentin the presently disclosed compositions will range from about 50% to100% of the amount normally present in a composition comprising thatagent as the only therapeutically active agent.

Compound 1 or pharmaceutical compositions thereof may also beincorporated into compositions for coating an implantable medicaldevice, such as prostheses, artificial valves, vascular grafts, stentsand catheters. Vascular stents, for example, have been used to overcomerestenosis (re-narrowing of the vessel wall after injury). However,patients using stents or other implantable devices risk clot formationor platelet activation. These unwanted effects may be prevented ormitigated by pre-coating the device with a pharmaceutically acceptablecomposition comprising a kinase inhibitor. Implantable devices coatedwith Compound 1 are another embodiment of the present invention.

All features of each of the aspects of the invention apply to all otheraspects mutatis mutandis.

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

Exemplification

As depicted in the Examples below, in certain exemplary embodiments,compounds are prepared according to the following general procedures. Itwill be appreciated that, although the general methods depict thesynthesis of certain compounds of the present invention, the followinggeneral methods, and other methods known to one of ordinary skill in theart, can be applied to all compounds and subclasses and species of eachof these compounds, as described herein.

Preparation of Compound 1

The synthesis of Compound 1 is described in detail at Example 3 of the'061 application.

Step 1:

In a 25 mL 3-neck round-bottom flask previously equipped with a magneticstirrer, Thermo pocket and CaCl₂ guard tube, N-Boc-1,3-diaminobenzene(0.96 g) and n-butanol (9.00 mL) were charged. The reaction mixture wascooled to 0° C. 2,4-Dichloro-5-trifluoromethylpyrimidine (1.0 g) wasadded dropwise to the above reaction mixture at 0° C.Diisopropylethylamine (DIPEA) (0.96 mL) was dropwise added to the abovereaction mixture at 0° C. and the reaction mixture was stirred for 1 hrat 0° C. to 5° C. Finally, the reaction mixture was allowed to warm toroom temperature. The reaction mixture was stirred for another 4 hrs atroom temperature. Completion of reaction was monitored by TLC usinghexane:ethyl acetate (7:3). The solid precipitated out was filtered offand washed with 1-butanol (2 mL). The solid was dried under reducedpressure at 40° C. for 1 hr. ¹H-NMR (DMSO-d6, 400 MHz) δ 1.48 (S, 9H),7.02 (m, 1H), 7.26 (m, 2H), 7.58 (S, 1H), 8.57 (S, 1H), 9.48 (S, 1H),9.55 (S, 1H).

Step 2:

To the above crude (3.1 g) in dichloromethane (DCM) (25 mL) was addedtrifluoroacetic acid (TFA) (12.4 mL) slowly at 0° C. The reactionmixture was allowed to warm to room temperature. The reaction mixturewas stirred for another 10 min at room temperature. The crude wasconcentrated under reduced pressure.

Step 3:

The concentrated crude was dissolved in DIPEA (2.0 mL) anddichloromethane (25 mL), and then cooled to −30° C. To the reactionmixture was slowly added acryloyl chloride (0.76 g) at −30° C. Thereaction mass was warmed to room temperature stirred at room temperaturefor 1.0 hr. The reaction was monitored on TLC using hexane:ethyl acetate(7:3) as mobile phase. The reaction was completed after 1 hr. Step 3yielded intermediate 1.

Step 4:

To obtain a salt of compound 1, a mixture of intermediate 1 (16 mg) and2-methoxy-4-(4-acetylpiperazinyl)aniline in dioxane (1.0 mL) withcatalytic trifluoroacetic acid was stirred overnight at 50° C. The crudewas concentrated under reduced pressure and purified using HPLC (TFAmodifier) to give compound 1 as a TFA salt. ¹H-NMR (DMSO-d₆, 400 MHz) δ10.2 (S, 1H), 8.2 (br, 1H), 8.30 (S, 1H), 7.73 (br, 1H), 7.52 (d, J=7.8Hz, 1H), 7.45 (d, J=7.8 Hz, 1H), 7.26 (J=8.2 Hz, 1H), 7.14 (be, 1H),6.60 (S, 1H), 6.42 (dd, J=11.4, 16.9 Hz, 1H), 6.24 (d, J=16.9 Hz, 1H),5.75 (d, J=11.4 Hz, 1H), 3.76 (S, 3H), 3.04 (br, 4H), 2.04 (S, 3H);calculated mass for C₂₇H₂₈F₃N₇O₃: 555.2. found: 556.2 (M+H⁺).

Step 5:

To obtain the free base form of Compound 1 from the TFA salt, the saltwas added to DCM and cooled to 0° C. Na₂CO₃ solution (9.6% w/w) wasadded at 0° C. The mixture was warmed to 20° C. and stirred for 35 min.The pH of the aqueous layer was >8. The layers were separated.Extraction of the aqueous layer was performed using DCM. The organiclayers were combined and washed with brine. The organic layer wascollected and evaporated to yield a solid of Compound 1.

General Procedures

X-ray powder diffraction (XRPD) analysis was carried out on a SiemensD5000, scanning the samples between 3 and 30 or 50° 2-theta. For samples<100 mg, ca. 5-10 mg of sample was gently compressed onto a glass slidewhich fitted into the sample holder. For samples >100 mg, ca. 100 mg ofsample was gently compressed into a plastic sample holder, so that thesample surface was smooth and just above the level of the sample holder.Measurements were made as follows:

step size 0.02 °2-theta scan step time 1 s offset 0 °2-theta divergenceslit type fixed divergence slit size 2.0000° receiving slit size 0.2 mmtemperature 20° C. anode material copper K-Alpha1 1.54060 AngstromsK-Alpha2 1.54443 Angstroms K-Beta 1.39225 Angstroms K-A2/K-A1 Ratio0.50000 Geneator settings 40 mA, 40 kV goniometer radius 217.50

In polarized light microscopy (PLM), the presence of crystallinity(birefringence) was determined using an Olympus BX50 polarisingmicroscope, equipped with a Motic camera and image capture software(Motic Images Plus 2.0). All images were recorded using the 20×objective, unless otherwise stated.

For thermogravimetric analysis (TGA), approximately 5-10 mg of materialwas accurately weighed into an open aluminium pan and loaded into asimultaneous thermogravimetric/differential thermal analyser (TG/DTA)and held at room temperature. The sample was then heated at a rate of10° C./min from 25° C. to 300° C. during which time the change in sampleweight was recorded along with any differential thermal events (DTA).Nitrogen was used as the purge gas, at a flow rate of 100 cm³/min.

For differential scanning calorimetry (DSC), approximately 5-10 mg ofmaterial was weighed into an aluminium DSC pan and sealednon-hermetically with a pierced aluminium lid. The sample pan was thenloaded into a Seiko DSC6200 (equipped with a cooler) cooled and held at25° C. Once a stable heat-flow response was obtained, the sample andreference were heated to ca. 260° C. at scan rate of 10° C./min and theresulting heat flow response monitored.

¹H-NMR experiments were performed on a Bruker AV400 (¹H frequency: 400MHz). ¹H experiments of each sample were performed in deuterated DMSOand each sample was prepared to ca. 10 mg concentration.

For dynamic vapour sorption (DVS), approximately 10 mg of sample wasplaced into a wire mesh vapour sorption balance pan and loaded into aDVS-1 dynamic vapour sorption balance by Surface Measurement Systems.The sample was subjected to a ramping profile from 20-90% relativehumidity (RH) at 10% increments, maintaining the sample at each stepuntil a stable weight had been achieved (99.5% step completion). Aftercompletion of the sorption cycle, the sample was dried using the sameprocedure, but all the way down to 0% RH and finally taken back to thestarting point of 20% RH. The weight change during thesorption/desorption cycles were plotted, allowing for the hygroscopicnature of the sample to be determined.

Infrared spectroscopy (IR) was carried out on a Bruker ALPHA Pspectrometer. Sufficient material was placed onto the centre of theplate of the spectrometer and the spectra were obtained using thefollowing parameters: resolution—4 cm⁻¹, background scan time—16 scans,sample scan time—16 scans, data collection 4000 to 400 cm⁻¹, resultspectrum—transmittance.

For Karl Fischer (KF) Coulometric titration, 10-15 mg of solid materialwas accurately weighed into a vial. The solid was then manuallyintroduced into the titration cell of a Mettler Toledo C30 CompactTitrator. The vial was back-weighed after the addition of the solid andthe weight of the added solid entered on the instrument. Titration wasinitiated once the sample had fully dissolved in the cell. The watercontent was calculated automatically by the instrument as a percentageand the data printed.

Reverse-phase gradient high performance liquid chromatography (HPLC) wasperformed on an Agilent 1100 instrument fitted with a C18, 3.0×100mm×3.5 μm column. The detection wavelength was 240 nm.

A Sotax AT7 dissolution bath (USP 2, EP 2 apparatus) was used for thedissolution study in which paddles were used to stir the media. Alltests were carried out at 37° C. and a paddle speed of 100 rpm.

Samples of each form were exposed to an environment of 40° C./75% RH for1 week and 2 week periods to determine stability. Resulting solids wereanalysed by XRPD and HPLC to establish if any changes had occurred.

Slurries of all each polymorphic form were created in deionised waterand shaken for ca. 24 hours. The resulting solid was then analysed byHPLC to determine the concentration of material dissolved.

Example 1 Preparation of Form A

ca. 120 mg of Compound 1 was weighed into a vial and slurried in ca. 2ml of acetonitrile. This was temperature cycled between ca. 0° C. andambient (ca. 22° C.) whilst stirring in 2 hour cycles for a period of2-3 days. Overnight, the sample was kept at ca. 2-5° C. Solid materialwas isolated and left to dry under vacuum for 7 days.

XRPD analysis (FIG. 1) showed the material to be crystalline. PLManalysis (not shown) indicated very fine, birefringent needle-likecrystals. TGA/DTA (FIG. 2) showed a 0.4% weight loss from the outset toca. 150° C. likely due to unbound solvent. No significant weight lossesseen prior to degradation. DSC analysis (FIG. 3) showed a singleendotherm at onset ca. 203.2° C. (peak 207.5° C.) due to the melt. IRanalysis (FIG. 4) corresponds with the input freebase material. ¹H NMR(not shown) carried out in deuterated DMSO showed a spectrum whichcorresponded with the input freebase. Acetonitrile does not appear to bepresent. DVS analysis (FIG. 5) showed a water uptake of 0.87% between 20and 70% RH, indicating a non-hygroscopic material. Post DVS XRPDindicated that the material remained Form A (data not shown). Nopolymorphic form changes were evident. Some loss in crystallinity wasobserved. KF analysis did not detect the presence of water. HPLC purityanalysis indicated a purity of ca. 97.6%. Form A could not be detectedby HPLC analysis for the aqueous solubility. The aqueous solubility istherefore poor.

XRPD analysis after 1 week storage (open container) at 40° C./75% RHshowed the material to still be consistent with Form A, with some lossin crystallinity. HPLC analysis indicated a purity of ca. 97.5%. XRPDanalysis after 2 week storage (open container) at 40° C./75% RH showedthe material to still be consistent with Form A, however, it becamepoorly crystalline. HPLC analysis indicated a purity of ca. 96.3%.

Example 2 Preparation of Form B

ca. 120 mg of Compound 1 was weighed into a vial and slurried in ca. 2ml tetrahydrofuran. This was temperature cycled between ca. 0° C. andambient (ca. 22° C.) whilst stirring in 2 hour cycles for a period of2-3 days. Overnight, the sample was kept at ca. 2-5° C. Solid materialwas isolated and left to dry under vacuum at ambient for 7 days and at40° C. for a further 2 days.

XRPD analysis (FIG. 6) showed the material produced to be crystalline.PLM analysis (not shown) indicated birefringent, rod-like crystals.TGA/DTA (FIG. 7) showed no significant weight losses prior todegradation after drying for 7 days at ambient under vacuum, and afurther 2 days at 40° C. DSC analysis (FIG. 8) showed an endotherm atonset 153.6° C. (peak 157.6°) directly followed by an exotherm at peak161.3° C. indicating a polymorphic transition. A further small endothermis present at peak 186.0° C., followed by a final endotherm at onset203.9° C. (peak 207.9° C.) which appears to correspond with the Form Amelt. IR analysis (FIG. 9) showed a significant number of differencesand shifts in comparison to Form A. ¹H NMR (not shown) carried out indeuterated DMSO showed a spectrum which corresponded with the inputfreebase. Traces of THF appear to be present. DVS analysis (FIG. 10)showed a water uptake of 0.74% between 20 and 70% RH, indicating anon-hygroscopic material. Post DVS XRPD (not shown) indicated that thematerial remained Form B. No polymorphic form changes were evident. KF(not shown) analysis indicated the presence of ca. 0.97% water. HPLCpurity analysis indicated a purity of ca. 97.0%. Form B could not bedetected by HPLC analysis for the aqueous solubility. The aqueoussolubility is therefore poor.

XRPD analysis after 1 week storage (open container) at 40° C./75% RHshowed the material to be predominantly amorphous. HPLC analysisindicated a purity of ca. 96.8%. XRPD analysis after 2 week storage(open container) at 40° C./75% RH showed the material to bepredominantly amorphous. HPLC analysis indicated a purity of ca. 95.9%.

In order to determine the relationship between Forms A and B, a sampleof Form B was heated to 120° C. and then XRPD analysis was carried out.The XRPD showed a mixture of Form B and Form A, indicating that as FormB is heated, it starts to convert to Form A. Form B was also heated to160° C. (temperature above phase transition seen in DSC) and then XRPDanalysis was carried out which indicated that the material convertedpredominantly to Form A at this temperature. The comparative XRPDresults are shown in FIG. 11.

Example 3 Preparation of Form C

ca. 120 mg of Compound 1 was weighed into a vial and slurried in ca. 100μl of DMF. This was temperature cycled between ca. 0° C. and ambient(ca. 22° C.) whilst stirring in 2 hour cycles. After ca. 2 hours, afurther 300 μl of DMF was added. The temperature cycling was continuedfor a period of 2-3 days. Overnight, the sample was kept at ca. 2-5° C.Solid material was isolated and left to dry under vacuum at ambient for7 days and at 40° C. for a further 2 days.

XRPD analysis (FIG. 12) showed the material to be crystalline. PLManalysis (not shown) indicated birefringent, rod-like crystals. TGA/DTA(FIG. 13) showed a weight loss of ca. 8.2%. (11.6 wt % DMF required fora mono solvate) after drying for 7 days at ambient under vacuum and afurther 2 days at 40° C. DSC analysis (FIG. 14) showed a broad endothermbetween 85-125° C., corresponding with the weight loss in the TGA. Afinal endotherm is seen at onset ca. 204.4° C. (peak 208.3° C.)corresponding with the Form A melt. IR analysis (FIG. 15) showed somedifferences and shifts in comparison with Form A. ¹H NMR (not shown)carried out in deuterated DMSO showed a spectrum which corresponds withthe input freebase with some DMF present (ca. 3:1 API:DMF). DVS analysis(FIG. 16) corresponded with the TGA data where the material is seen tocontain solvent, which is lost as the relative humidity is increased.Post DVS XRPD (not shown) indicated that the material converted to FormA during DVS analysis. KF analysis (not shown) indicated the presence ofca. 0.01% water. HPLC purity analysis indicated a purity of ca. 97.3%.Form C could not be detected by HPLC analysis for the aqueoussolubility. The solubility is therefore poor.

XRPD analysis after 1 week storage (open container) at 40° C./75% RHshowed the material converted to Form A with some loss in crystallinity.HPLC analysis indicated a purity of ca. 97.1%. XRPD analysis after 2week storage (open container) at 40° C./75% RH showed the material to bepoorly crystalline with the peaks present corresponding with Form A.HPLC analysis indicated a purity of ca. 96.8%. The comparative XRPDresults are shown in FIG. 17.

To determine whether Form C remains the same after losing the solventpresent, a sample of Form C was heated to 120° C. (i.e., just past thetemperature at which the solvent is removed) and then XRPD analysis wascarried out. The XRPD showed that the material had converted to Form A.Similarly, after drying at ambient temperature under vacuum for 7 daysand then for a further 2 days at 40° C., Form C showed some loss incrystallinity and converted to Form A. The comparative XRPD results areshown in FIG. 18.

Example 4 Preparation of Form D

ca. 120 mg of Compound 1 was weighed into a vial and slurried in ca. 2ml of 1,4-dioxane. This was temperature cycled between ca. 0° C. andambient (ca. 22° C.) whilst stirring in 2 hour cycles for a period of2-3 days. Overnight, the sample was kept at ca. 2-5° C. Solid materialwas isolated and left to dry under vacuum at ambient for 7 days and at40° C. for a further 2 days.

XRPD analysis showed that the preparation of Form D provided aboveresulted in a mixture of Form B and Form D (FIG. 19). PLM analysisindicated birefringent, rod-like crystals (not shown). TGA/DTA (FIG.20A) showed a weight loss of ca. 5.5% between 60-100° C. after dryingfor 7 days at ambient under vacuum. After drying for 7 days at ambientunder vacuum, and a further 2 days at 40° C., the TGA (FIG. 20B) showeda weight loss of ca. 1.4% between 100-150° C. DSC analysis after 40° C.drying (FIG. 21) showed a very small exotherm at ca. 146° C. A finalendotherm was then seen at onset ca. 202.6° C. (peak 207.4° C.)corresponding with the Form A melt. IR analysis (FIG. 22) correspondedwith the spectrum of Form B, with small 1,4-dioxane peaks present.¹H-NMR (not shown) carried out in deuterated DMSO after 7 days of dryingat ambient temperature under vacuum showed a spectrum which correspondswith the input free base with some 1,4-dioxane present (ca. 2:1 API:1,4-Dioxane). DVS analysis (FIG. 23) corresponded with the TGA datawhere the material is seen to contain solvent which is lost as therelative humidity is increased. Post DVS XRPD (not shown) indicated thatthe material converted completely to Form B during DVS analysis. KFanalysis (not shown) indicated the presence of ca. 1.1% water. HPLCpurity analysis indicated a purity of ca. 96.6%. Form D could not bedetected by HPLC analysis for the aqueous solubility. The solubility istherefore extremely poor.

XRPD analysis after 1 week storage (open container) at 40° C./75% RHshowed the material converted completely to Form B with some loss incrystallinity. HPLC analysis indicated a purity of ca. 96.5%. XRPDanalysis after 2 week storage (open container) at 40° C./75% RH showedthe material to be poorly crystalline with the peaks presentcorresponding with Form B. HPLC analysis indicated a purity of ca.95.5%. The comparative XRPD results are shown in FIG. 24.

Example 5 Preparation of Form E

ca. 120 mg of Compound 1 was weighed into a vial and ca. 12 ml of MEKwas then added in attempts to dissolve the material. A very thin slurryresulted and this was then filtered to obtain a saturated solution. Thesolution was placed at ca.-18° C. for crash cooling for 2-3 days. Solidmaterial was isolated and left to dry under vacuum at ambient for 7 daysand at 40° C. for a further 2 days.

XRPD analysis (FIG. 25) showed the material to be crystalline. PLManalysis (not shown) indicated birefringent, thin, rod-like crystals.TGA/DTA (FIG. 26) showed a weight loss of ca. 5.2% between ca. 70-110°C. after drying for 7 days at ambient under vacuum. DSC analysis (FIG.27) showed a broad endotherm between 70-110° C., corresponding with theweight loss in the TGA. A final endotherm is seen at onset ca. 198.4° C.(peak 203.3° C.). IR analysis (FIG. 28) showed some small shifts incomparison with Form A. ¹H-NMR (not shown) carried out in deuteratedDMSO after 7 days of drying at ambient under vacuum showed a spectrumwhich corresponds with the input free base with a non-stoichiometricamount of MEK present. DVS analysis (FIG. 29) showed a water uptake of0.25% between 20 and 70% RH. Post DVS XRPD (not shown) indicated thatthe material converted to Form A during DVS analysis. KF analysis (notshown) indicated the presence of ca. 1.2% water. HPLC purity analysisindicated a purity of ca. 97.8%. Form E could not be detected by HPLCanalysis for the aqueous solubility. The solubility is thereforeextremely poor.

XRPD analysis after 1 week storage (open container) at 40° C./75% RHshowed the material to be predominantly amorphous with visible peakspresent corresponding with Form A. HPLC analysis indicated a purity ofca. 97.7%. XRPD analysis after 2 week storage (open container) at 40°C./75% RH showed the material to be predominantly amorphous with visiblepeaks present corresponding with Form A. HPLC analysis indicated apurity of ca. 97.3%. The comparative XRPD results are shown in FIG. 30.

In order to determine whether Form E remains the same after losing thesolvent present, a sample of Form E was heated to 120° C. (i.e., justpast the temperature at which the solvent is lost) and then XRPDanalysis was carried out. The XRPD showed that the material hadconverted to Form A. Similarly, after drying at ambient under vacuum for7 days and then for a further 2 days at 40° C., Form E lost somecrystallinity and converted to Form A. The comparative XRPD results areshown in FIG. 31.

Example 6 Preparation of Form F

ca. 120 mg of Compound 1 was weighed into a vial and ca. 200 μl of NMPwas then added in order to dissolve the material. Anti-solvent additionwas then carried out by adding a total of ca. 4.5 ml of TBME in 500 μlaliquots to obtain a cloudy solution. The sample was then allowed tostand for ca. 2 hours before the solid was isolated. The solid materialwas then left to dry under vacuum at ambient for 7 days and at 40° C.for a further 2 days.

XRPD analysis (FIG. 32) showed the material to be crystalline. PLManalysis (not shown) indicated birefringent, very thin, plate-likecrystals which appear to grow in clusters. TGA/DTA (FIG. 33) showed aweight loss of ca. 10.2% between 70-120° C. (15.1 wt % NMP required fora mono solvate) after drying for 7 days at ambient temperature undervacuum. DSC analysis (FIG. 34) showed an endotherm at onset 97° C. (peak100.9° C.) corresponding with the weight loss in the TGA. A finalendotherm is seen at onset ca. 204.2° C. (peak 208.3° C.) correspondingwith the Form A melt. IR analysis (FIG. 35) showed some small shifts incomparison with Form A. ¹H-NMR (not shown) carried out in deuteratedDMSO after 7 days of drying at ambient under vacuum showed a spectrumwhich corresponds with the input free base with a non-stoichiometricamount of NMP present. DVS analysis (FIG. 36) corresponded with the TGAdata where the material is seen to contain solvent, which is lost as therelative humidity is increased. Post DVS XRPD (not shown) indicated thatthe material converted predominantly to Form A (traces of Form Fremaining). KF analysis (not shown) indicated the presence of ca. 0.87%water. HPLC purity analysis indicated a purity of ca. 97.0%. Form Fcould not be detected by HPLC analysis for the aqueous solubility. Thesolubility is therefore poor.

XRPD analysis after 1 week storage (open container) at 40° C./75% RHshowed the material converted to Form A, with some loss incrystallinity. HPLC analysis indicated a purity of ca. 96.8%. XRPDanalysis after 2 week storage (open container) at 40° C./75% RH showedthe material to be partially crystalline with peaks presentcorresponding with Form A. HPLC analysis indicated a purity of ca.96.3%. The comparative XRPD results are shown in FIG. 37.

In order to determine whether Form F remains the same after losing thesolvent present, a sample of Form F was heated to 120° C. (i.e., justpast the temperature at which the solvent is lost) and then XRPDanalysis was carried out. The XRPD showed that the material hadconverted predominantly to Form A. Similarly, after drying at ambienttemperature under vacuum for 7 days and then for a further 2 days at 40°C., Form F lost some crystallinity and converted predominantly to FormA. The comparative XRPD results are shown in FIG. 38.

Example 7 Preparation of Form G

ca. 120 mg of Compound 1 was weighed into a vial and slurried in ca. 100μl of NMP. This was temperature cycled between ca. 0° C. and ambient(ca. 22° C.) whilst stirring in 2 hour cycles for a period of 2-3 days.Overnight, the sample was kept at ca. 2-5° C. Solid material wasisolated and left to dry under vacuum at ambient for 7 days and at 40°C. for a further 2 days. A portion of the solid was also dried at 80° C.for ca. 4 days.

XRPD analysis (FIG. 39) showed the material to be crystalline. PLManalysis (not shown) indicated birefringent, plate-like crystals.TGA/DTA (FIG. 40A) showed weight losses of ca. 23.6% and 10.2% afterdrying for 4 days at ambient temperature under vacuum. After drying for7 days at ambient temperature under vacuum and a further 2 days at 40°C., the TGA (FIG. 40B) showed a weight loss of ca. 6.3% between ca.70-110° C. After drying for 4 days at 80° C., the TGA (FIG. 40C) showeda weight loss of ca. 2.8% between ca. 70-110° C. DSC analysis afterdrying at 80° C. (FIG. 41) showed an endotherm at onset ca. 205.3° C.(peak 210.0° C.). IR analysis after 4 days of ambient temperature drying(FIG. 42) showed some shifts in comparison with Form A and also thepresence of NMP. ¹H-NMR (not shown) carried out in deuterated DMSO after4 days of drying at ambient temperature under vacuum corresponded withthe input free base with a significant amount of NMP present. DVSanalysis (FIG. 43) corresponded with the TGA data, where the material isseen to contain significant amounts of solvent that are lost as therelative humidity is increased. Post DVS XRPD (not shown) indicated thatthe material converted to Form A. KF analysis (not shown) indicated thepresence of ca. 0.45% water. HPLC purity analysis indicated a purity ofca. 97.0%. Form F in the HPLC chromatogram to determine aqueoussolubility was below the limit of quantification. The aqueous solubilityis therefore poor.

XRPD analysis after 1 week storage (open container) at 40° C./75% RHshowed the material converted predominantly to Form A, with some loss incrystallinity. HPLC analysis indicated a purity of ca. 96.8%. XRPDanalysis after 2 week storage (open container) at 40° C./75% RH showedthe material converted predominantly to Form A, with some loss incrystallinity. HPLC analysis indicated a purity of ca. 96.2%. Thecomparative XRPD results are shown in FIG. 44.

After drying at ambient under vacuum for 7 days and then for a further 2days at 40° C., Form G converted to Form A. After drying at 80° C. for 4days, Form G converted to Form A. The comparative XRPD results are shownin FIG. 45.

Example 8 Polymorph Stability Studies

The results from the competitive slurries carried out at ambient (ca.22° C.) and 60° C. are tabulated below.

TABLE 1 Competitive Slurry results at ambient and 60° C. Solvent SystemAmbient (approx. 22° C.) 60° C. Dichloromethane Form B Form BIsopropanol Form A Form A Acetone Form A Form A Ethyl Acetate Form AForm A Acetone:Water (80:20) Form A Form HFrom slurrying at both ambient and 60° C., the majority of experimentsresulted in conversion to Form A, thus indicating that Form A is likelythe more stable form in comparison with Form B. Form B has been obtainedconsistently throughout the study from dichloromethane andtetrahydrofuran. A new polymorphic form, labelled Form H, was obtainedfrom the competitive slurry carried out in acetone:water (80:20) at 60°C.

Example 9 Preparation of Form H

ca. 10 mg of Form A and ca. 10 mg of Form B were weighed into a vial.ca. 200 μl of acetone:water (80:20%) was added to the vial to form aslurry. The sample was allowed to stir at ca. 50° C. for 3 days. Thesolid material was isolated and left to dry at ambient before analysiswas carried out. Further drying was also carried out at 40° C. for 2days.

XRPD analysis (FIG. 46) showed the material to be crystalline, with thediffractogram different from all identified polymorphic forms. PLManalysis (not shown) indicated birefringent, block-like crystals. Thematerial visually appeared yellow in colour. TGA/DTA (FIG. 47) after 2days of drying at 40° C., showed a 4.4% weight loss from the outset upto ca. 120° C. The final endotherm in the DTA trace appears tocorrespond with the Form A melt. (3.14 wt % is required for 1 moleequivalent of water.)

To examine whether Form H changes to Form A after heating to 115° C.(point after the solvent loss), the Form H material was heated to 115°C. and XRPD analysis and DSC analysis were then carried out. The XRPDdiffractogram (FIG. 48) after heating still corresponded with Form H,with some loss in crystallinity likely due to the harsh heatingconditions. The DSC analysis (FIG. 49) indicated overlapping endothermsbetween ca. 115-135° C., followed by an exotherm at peak 143.9° C.,likely indicating a polymorphic transition. A final endotherm waspresent at onset 202.7° C. (peak 206.6° C.) corresponding with the FormA melt.

To test whether Form H picks up water after desolvating/dehydrating thematerial by heating to 115° C., a further test was carried out wherebyForm H was heated to 115° C. in a TGA pan. The sample was then removedfrom the TGA pan and allowed to sit on the bench for ca. 1 hour. After 1hour, another TGA was carried out up to 115° C. TGA for the sampleheated to 115° C. showed a ca. 4.2% weight loss of the solvent/waterpresent (FIG. 50). TGA after leaving the desolvated/dehydrated materialon the bench for ca. 1 hour, showed a ca. 3.7% loss up to 115° C. (FIG.51). The material therefore picked up water upon standing at ambientconditions on the bench. This indicates that it is either hygroscopic orrapidly rehydrates following dehydration/desolvation.

Example 10 Preparation of Form I

Approximately ca. 5 mL of acetonitrile:water (10%) was added to ca. 1 gof Compound 1 free base to form a slurry. In a separate vial, ca. 3 mLof acetonitrile:water (10%) was added to 1 equivalent of hydrobromicacid (48%). The acid solution was then added dropwise over a 1 hourperiod to the free base slurry whilst stirring and maintaining atemperature between 0-5° C. After the complete addition of the acid, afurther 3 mL of acetonitrile:water (10%) was added. The reaction wasstirred for ca. 1 day before being isolated and dried under vacuum atambient (ca. 22° C.). A yield of ca. 79% was obtained.

Compound 1 hydrobromide salt material was ground using a Retsch BallMill for ca. 25 minutes, with a 5 minute break midway to prevent thesample from overheating.

Approximately 500 mg of amorphous Compound 1 hydrobromide salt materialwas slurried in ca. 18 mL of acetone:water (90:10). The suspension wasthen temperature cycled between 4 and 25° C. in four hour cycles for ca.2 days, before being isolated and dried under vacuum at ambient (ca. 22°C.). The secondary screen analysis was carried out on Form I afterdrying.

Form I was scaled-up for further analysis. During the scale-up, a colourchange was observed from yellow to a light beige/cream colour. XRPDanalysis (FIG. 52) showed the material produced from scale-up to becrystalline and predominantly consistent with the small scale Form Idiffractogram. IR and ¹H NMR are depicted in FIG. 53 and FIG. 54,respectively. PLM analysis indicated birefringent, fibrous, needle-likecrystals when wet. Upon drying the material appeared to lose itsneedle-like morphology, appearing as small particles with no clearlydefined morphology. Hot stage microscopy indicated melting at ca. 135°C. with some recrystallization occurring at ca. 180° C., followed bycomplete melting by ca. 210° C. After drying under vacuum for ca. 72hours, the TGA/DTA indicated a weight loss of 2.8% from ca. 80 to 120°C. corresponding with an endotherm in the DTA trace (FIG. 55). Anexotherm was observed in the DTA trace at onset ca. 149° C. (peak ca.166° C.), followed by a further endotherm at onset ca. 197° C. (peak ca.201° C.). After standing at ambient conditions, the TGA/DTA was re-runshowing a weight loss of 2.6% from the outset to ca. 80° C., followed bya further weight loss of 2.8% between ca. 80° C. and 120° C.corresponding with two endotherms in the DTA trace (FIG. 56). Anexotherm was then observed in the DTA trace at onset ca. 155° C. (peakca. 167° C.) followed by a further endotherm at onset ca. 196° C. (peakca. 202° C.). The DSC analysis indicated overlapping endotherms startingfrom the outset, followed by an exotherm at onset ca. 138° C. (peak ca.149° C.) and a further endotherm at onset ca. 92° C. (peak ca. 200° C.)(FIG. 57). DVS analysis (FIG. 58) showed the following observations:

-   -   Cycle 1—Sorption 20-90% RH    -   Sample gradually takes up ca. 0.66% mass.    -   Cycle 2—Desorption 90-0% RH    -   Between 90-10% RH, sample mass decreases gradually by ca. 1.2%.    -   A rapid loss of ca. 2.7% occurs between 10-0% RH.    -   Cycle 3—Sorption 0-20% RH    -   Moisture uptake of ca. 2.8% between 0-20% RH.

The input material containing ca. 5.6% water appeared to be relativelynon-hygroscopic. Approximately 1 equivalent of water was lost at thelower RH percentages. Post DVS XRPD analysis indicated that the materialremained as Form I (FIG. 59). No polymorphic form changes were evident.KF analysis indicated the presence of ca. 5.4% water. HPLC purityanalysis indicated a purity of ca. 99.66% (FIG. 60). Ion chromatographyindicated the presence of 1.64% bromide (ca. 12.57% required for 1equivalent). XRPD analysis carried out on the thermodynamic solubilityexperiment solids remaining after 24 hours, indicated that for pH 6.6,4.5 and 3.0 the material remained as Form I (FIG. 61). For pH 1, thematerial appeared to be a mixture of Form I and possibly the HCl saltformed during screening.

From the characterisation carried out on Form I, this form wasdetermined to be a hydrated version of the freebase rather than abromide salt form. The TGA/DTA and DVS data appear to suggest that thismay be either a hygroscopic monohydrate or a dihydrate form.

7 Day Stability Studies at 25° C., 80° C., 40° C./75% RH (Open andClosed Conditions).

Approximately 15 mg of Form I was placed separately into vials and thenexposed to 25° C., 80° C. and 40° C./75% RH environments (open andclosed vials) for 1 week to determine stability. The resulting solidswere analysed by XRPD and HPLC to establish if any changes had occurred.Results are presented in Tables 2 and 3 and FIGS. 62 and 63.

TABLE 2 1 week stability studies (Open container) Condition Purity XRPDanalysis 40° C./75% RH 98.9% Form I 80° C. 98.5% Form I (some loss incrystallinity) 25° C. 98.7% Form I

TABLE 3 1 week stability studies (Closed container) Condition PurityXRPD Analysis 40° C./75% RH 99.3% Form I (some loss in crystallinity)80° C. 99.2% Form I (some loss in crystallinity) 25° C. 99.4% Form I

Thermodynamic Solubility Studies.

Slurries of Form I were created in media of various pH (pH 1; pH 3; pH4.5 and pH 6.6) and shaken for ca. 24 hours. After 24 hours, theslurries were filtered and the solution analysed by HPLC in order todetermine the solubility at the various pH levels. For the buffersolutions, KCl/HCl was used for pH 1 and citrate/phosphate combinationsfor pH 3, 4.5 and 6.6 (10 mM). The pH of the solutions was also measuredprior to HPLC analysis. XRPD analysis was carried out on the remainingsolids after 24 hours of shaking. Results are presented in Table 4:

TABLE 4 Thermodynamic Solubility studies Buffer pH pH prior to analysisSolubility (mg/mL) 1 0.95 3.266 3 2.26 0.023 4.5 3.38 0.002 6.6 5.04 Notdetected

We claim:
 1. A solid form of Compound 1:

wherein Compound 1 is a free base.
 2. The solid form of claim 1, whereinthe solid form is amorphous.
 3. The solid form of claim 1, wherein thesolid form is crystalline.
 4. The solid form of claim 3, whereinCompound 1 is unsolvated.
 5. The solid form of claim 4, wherein thesolid form is Form A.
 6. The solid form of claim 5, having one or morepeaks in its X-ray powder diffraction pattern selected from those atabout 6.73, about 18.30, about 18.96 and about 25.48 degrees 2-theta. 7.The solid form of claim 4, wherein the solid form is Form B.
 8. Thesolid form of claim 7, having one or more peaks in its X-ray powderdiffraction pattern selected from those at about 10.67, about 12.21,about 18.11, about 19.24 and about 21.53 degrees 2-theta.
 9. The solidform of claim 3, wherein Compound 1 is a dimethylformamide solvate. 10.The solid form of claim 9, wherein the solid form is Form C.
 11. Thesolid form of claim 10, having one or more peaks in its X-ray powderdiffraction pattern selected from those at about 16.32, about 18.82,about 20.26, about 22.58 and about 25.36 degrees 2-theta.
 12. The solidform of claim 3, wherein Compound 1 is a 1,4-dioxane solvate.
 13. Thesolid form of claim 12, wherein the solid form is Form D.
 14. The solidform of claim 13, having one or more peaks in its X-ray powderdiffraction pattern selected from those at about 18.40, about 19.31,about 20.14, about 20.53 and about 25.25 degrees 2-theta.
 15. The solidform of claim 3, wherein Compound 1 is a methyl ethyl ketone solvate.16. The solid form of claim 15, wherein the solid form is Form E. 17.The solid form of claim 16, having one or more peaks in its X-ray powderdiffraction pattern selected from those at about 5.78, about 12.57,about 15.34, about 19.10 and about 24.80 degrees 2-theta.
 18. The solidform of claim 3, wherein Compound 1 is a N-methyl-2-pyrrolidone solvate.19. The solid form of claim 18, wherein the solid form is Form F. 20.The solid form of claim 19, having one or more peaks in its X-ray powderdiffraction pattern selected from those at about 15.51, about 16.86,about 18.80, about 20.97 and about 23.32 degrees 2-theta.
 21. The solidform of claim 18, wherein the solid form is Form G.
 22. The solid formof claim 21, having one or more peaks in its X-ray powder diffractionpattern selected from those at about 6.79, about 17.86, about 19.43,about 19.98 and about 22.35 degrees 2-theta.
 23. The solid form of claim3, wherein Compound 1 is a hydrate.
 24. The solid form of claim 23,wherein the solid form is Form H.
 25. The solid form of claim 24, havingone or more peaks in its X-ray powder diffraction pattern selected fromthose at about 10.82, about 11.08, about 18.45, about 22.85 and about25.06 degrees 2-theta.
 26. The solid form of claim 23, wherein the solidform is Form I.
 27. The solid form of claim 26, having one or more peaksin its X-ray powder diffraction pattern selected from those at about6.13, about 12.22, about 15.91, about 18.35, about 18.88, and about21.90 degrees 2-theta.
 28. A composition comprising the solid form ofclaim 1-and a pharmaceutically acceptable carrier or excipient.
 29. Amethod for inhibiting at least one mutant of EGFR selectively ascompared to wild type EGFR (WT EGFR), in a biological sample or in apatient, comprising contacting the biological sample with, oradministering to the patient, a solid form according to claim 1, or acomposition thereof.
 30. The method according to claim 29, wherein saidmethod is sparing for WT EGFR.
 31. The method according to claim 29,wherein the at least one mutant is an activating mutant, a deletionmutant, a point mutation, or a mutant selected from T790M, delE746-A750,L858R or G719S.
 32. A method for treating a mutant EGFR-mediateddisorder or condition in a patient, comprising administering to thepatient a composition according to claim
 28. 33. The method of claim 32,wherein the disorder or condition is a cancer.